Aromatic prenyltransferases, nucleic acids encoding same and uses therefor

ABSTRACT

In accordance with the present invention, a novel aromatic prenyltransferase, Orf2 from  Streptomyces  sp. strain CL190, involved in naphterpin biosynthesis has been identified and the structure thereof elucidated. This prenyltransferase catalyzes the formation of a C—C bond between a prenyl group and a compound containing an aromatic nucleus, and also displays C—O bond formation activity. Numerous crystallographic structures of the prenyltransferase have been solved and refined, e.g., (1) prenyltransferase complexed with a buffer molecule (TAPS), (2) prenyltransferase as a binary complex with geranyl diphosphate (GPP) and Mg 2+ , and prenyltransferase as ternary complexes with a non-hydrolyzable substrate analogue, geranyl S-thiolodiphosphate (GSPP) and either (3) 1,6-dihydroxynaphthalene (1,6-DHN), or (4) flaviolin (i.e., 2,5,7-trihydroxy-1,4-naphthoquinone, which is the oxidized product of 1,3,6,8-tetrahydroxynaphthalene (THN)). These structures have been solved and refined to 1.5 Å, 2.25 Å, 1.95 Å and 2.02 Å, respectively. This first structure of an aromatic prenyltransferase displays an unexpected and non-canonical (β/α)-barrel architecture. The complexes with both aromatic substrates and prenyl containing substrates and analogs delineate the active site and are consistent with a proposed electrophilic mechanism of prenyl group transfer. These structures also provide a mechanistic basis for understanding prenyl chain length determination and aromatic co-substrate recognition in this structurally unique family of aromatic prenyltransferases. This structural information is useful for predicting the aromatic prenyltransferase activity of proteins.

RELATED APPLICATIONS

This application is a divisional of non provisional application Ser. No.11/342,328 filed Jan. 27, 2006, which claims priority from U.S.provisional application No. 60/648,046, filed Jan. 28, 2005, the entirecontents of each of which are hereby incorporated by reference herein.

FIELD OF THE INVENTION

The present invention relates to aromatic prenyltransferases, nucleicacids encoding same, crystalline forms of aromatic prenyltransferases,and various uses therefor. In one embodiment, methods are provided forpredicting the activity and/or substrate specificity of putativearomatic prenyltransferases. In another embodiment, methods of screeningcompounds to identify compounds which bind aromatic prenyltransferasesand/or modulate the activity thereof, are provided. In yet anotherembodiment, methods of screening compounds to identify potentialsubstrates of aromatic prenyltransferases are provided. In still anotherembodiment, methods are provided for prenylating aromatic structures, aswell as controlling and/or modifying the degree of prenylation promotedby aromatic prenyltransferases. In a further embodiment, methods areprovided for identifying proteins having the newly discovered beta/alphabarrel structure. In a still further embodiment, methods are providedfor controlling and or modifying the substrate specificity of aromaticprenyltransferases.

BACKGROUND OF THE INVENTION

Nature is a prolific producer of small molecules that have evolved tointeract with diverse biological targets. From a human healthperspective, natural products have dramatically altered our lives byproviding many front-line drugs as well as chemical probes to unravelbasic molecular pathways germane to health and disease. Although naturalproducts continue to provide about half of all new chemical entitiesapproved as drugs by the U.S. Food and Drug Administration, drugdiscovery during the latter part of the 20^(th) century shifted awayfrom natural products towards synthetic libraries. This paradigm shiftreflected the complexity of small, natural libraries against thesimplicity of large, combinatorial synthetic libraries and wasrationalized in order to keep pace with the enormous capacity ofindustrial high-throughput screening programs. New drugs fromcombinatorial chemical libraries, however, did not materialize duringthis time period, while natural products continued as an importantsource. Natural products, like drugs, cover a chemical space that ismuch more diverse than combinatorial compounds, thereby reflecting therich chemical diversity of this resource.

Recent technological advances in natural product research involvingisolation, characterization, synthesis, and biosynthesis have rekindledan interest in their investigation in academia and industry. With theadvent of modern molecular biology, the field of biosynthesis hasblossomed over the past decade with new approaches to generatebiosynthetic libraries that further extend natural product structuraldiversity into new chemical space. In vivo approaches involvingcombinatorial biosynthesis, mutasynthesis, and precursor-directedbiosynthesis and complementary in vitro approaches that combine chemicalsynthesis and enzymology (chemoenzymatic synthesis) have led toimpressive libraries of novel molecules never encountered in nature.Natural product structural classes that have been biosyntheticallymanipulated in this fashion include the polyketides, nonribosomalpeptides, terpenoids, and alkaloids. Most progress in this burgeoningfield has resided with the actinomycetes (soil bacteria), which offerimpressive arrays of natural products whose biosynthetic genes aretypically clustered and are thus readily amenable to geneticmanipulation. One notable exception that is absent from the biosyntheticdiversification platform, however, is the hybrid isoprenoid class ofnatural products.

Natural products, such as the isoprenoid (terpenoid) family of diversechemical scaffolds have held significant interest for the syntheticorganic chemistry community because they are both challenging syntheticprojects and possess varied biological activities and medicinalproperties. Within the terpenoid family, the total synthesis ofsesquiterpene natural products and related analogs continue to dominatethe chemical literature. The demand for a reliable production platformfor structurally complex terpenes has increased dramatically over thelast 10 years and is of growing interest. Elegant synthetic schemes forterpenoids have been developed, but suffer from low yields and lowregio- and enantio-selectivity. Although engineered E. coli has thepotential to make mg/L levels of sesquiterpene hydrocarbons, the morebiologically active terpenes are highly functionalized with hydroxyl,methyl, acetyl, halide, carbohydrate, and peroxide functional groupsthat require multi-step biosynthetic mechanisms often tethered toendo-membrane systems conducive for metabolic coupling. By integratingbiosynthetic complexity with synthetic diversification, it may bepossible for many of these hurdles to the technological development ofterpenoids to be overcome.

Moreover, hybrid compounds containing terpene-derived residues comprisea large and diverse group of natural products that command an importantrole in human health (see Table 1). Historically this class of compoundshas provided important drugs (e.g., the anticancer agent vincristine,the antimalarial quinine and the immunosuppressant mycophenolatemofetil) as well as challenging synthetic targets (e.g., strychnine andreserpine). In addition to natural products, many important coenzymes(ubiquinone and plastoquinone) and vitamins (tocopherols,phylloquinones, and menaquinones), which function in electron transportsystems, contain isoprenoid residues.

TABLE 1 Representative hybrid isoprenoids, their sources and biologicalsignificance Natural Product Source Isoprenoid Hybrid BiologicalActivity mycophenolic fungus polyketide immunosuppressant acid khellinplant polyketide bronchial asthma tetrahydro- plant polyketide narcotic,antiemetic cannabinol rotenone plant isoflavonoid insecticide psoralenplant coumarin skin pigment and irritant novobiocin bacterium coumarinantibiotic lucidin plant quinine mutagen emetine plant tetrahydroiso-alkaloid emetic quinoline (ipecac) ergometrine fungus ergotalkaloidoxytocic reserpine plant indole alkaloidanti- hypertensivevincristine plant indole alkaloid anticancer strychnine plant indolealkaloid toxin lyngbyatoxin cyano- indole alkaloid inflammatory agentbacterium quinine plant quinoline alkaloid antimalarial camptothecinplant quinoline alkaloid topoisomerase/ inhibitor

Nature has assembled a myriad of scaffolds to which isoprenoids havebeen attached, and these include polyketides (the so-calledmeroterpenoids), flavonoids, coumarins, quinones, alkaloids, phenazines,and the like. Often the terpenoid unit is further elaborated byelectrophilic cyclization and oxidative chemistry upon attachment to itsbuilding block, thereby leading to the great structural diversityobserved within this group. While most of these natural products containa single isoprenoid unit of varying chain length, others harbor multipleisoprene units such as in the tetraprenylated benzoylphloroglucinolderivatives sampsoniones A-I.

The vast majority of hybrid isoprenoids are derived from eukaryotes,particularly plants. For instance, over a thousand monoterpenoid indolealkaloids have been characterized, making this a major class of plantalkaloids. On the other hand, terpenoids, and in particular hybridisoprenoids, appear to have a limited distribution in prokaryotes. Whileactinomycetes are metabolically very rich bacteria and produce manyimportant biosynthetic classes of natural products that includepolyketides, nonribosomal peptides, aminoglycosides, and the like, theterpenoids are notably scarce. As a consequence, while other naturalproduct structural classes have been biosynthetically exploited in thedrug discovery arena, the hybrid isoprenoids are noticeably absent dueto our limited understanding of their biosynthesis at the biochemicaland genetic levels.

The majority of the basic understanding of how hybrid isoprenoids arebiosynthesized in plants, fungi and bacteria is based on feedingexperiments with labeled precursors. Enzymes and their encoding genesassociated with interfacing isoprenyl diphosphates with their smallmolecule building blocks are very few and are mostly associated withplant natural products such as shikonin and with coenzymes and vitaminssuch as the ubiquinones, plastoquinones, menaquinones, and tocopherols.Very recently, two prokaryotic prenyltransferases (PTases) involved inthe biosynthesis of the streptomycete antibiotics clorobiocin andnovobiocin and the cyanobacterial toxin lyngbyatoxin were discovered.These soluble, monomeric PTases contrast with the membrane-associatedPTases previously identified from eukaryotes.

Actinomycetes produce a limited set of pure and hybrid terpenoids. Theantibiotic novobiocin was the first streptomycete natural productdiscovered with a terpenoid side chain; this group has since grown toinclude other members bearing naphthoquinones (naphterpin, furaquinocin,napyradiomycins), phenazines (lavanducyanin, aestivophoenin),shikimate-derived quinones, and other aromatic substrates (see FIG. 1B).Feeding experiments delineated a number of biosynthetic pathways,including those to novobiocin, naphterpin, and furaquinocin, andrevealed that actinomycetes utilize both the mevalonate andnonmevalonate (methyl-D-erythritol 4-phosphate (MEP)) pathways tosynthesize their isoprene building blocks.

The development of novel methodologies related to natural productschemistry and biosynthesis is of growing interest. Prenylated aromaticnatural products appear to be a very promising class of therapeuticallycompounds. The prenylation of aromatic compounds often leads tosignificant alteration in the bioactivity profile of a compound, by boththe creation of a novel C—C bond and also the introduction of one ormore double bonds in the framework of the final product. Such compoundscan affect a wide variety of biological systems in mammals and includeroles as anti oxidants, anti-inflammatories, anti-virals,anti-proliferatives, and anti-cancers.

Prenyltransferases (PTases) are ubiquitous enzymes that catalyze thealkylation of electron rich prenyl acceptors by the alkyl moieties ofallylic isoprene diphosphates. Prenyltransferases utilize isoprenoiddiphosphates as substrates, and catalyze the addition of the acyclicprenyl moiety to isopentenyl diphosphate (IPP), higher order prenyldiphosphates, aromatic rich molecules and proteins. Until now, only afew “aromatic” prenyltransferases have been isolated, each of which hasbeen shown to interact with only a limited range of substrate(s) and/orprenyl donors. Such prenyltransferases have otherwise only beennominally characterized; and none of such prenyltransferases have beencharacterized at the structural level.

Accordingly, there is a need in the art for the identification of novelenzymes capable of promoting the prenylation of aromatic compounds, aswell as compounds which can modulate the prenylation of aromaticcompounds. These and other needs are addressed by the present invention,as described in greater detail in the specification and claims whichfollow.

SUMMARY OF THE INVENTION

In accordance with the present invention, a novel aromaticprenyltransferase, Orf2 from Streptomyces sp. strain CL190, involved innaphterpin biosynthesis (Shin-ya, et al., in J. Antibiot. (Tokyo) 43,444-447 (1990)) has been identified and the structure thereofelucidated. This prenyltransferase catalyzes the formation of a C—C bondbetween a prenyl group and a compound containing an aromatic nucleus,and also displays C—O bond formation activity. Numerous crystallographicstructures of the prenyltransferase have been solved and refined, e.g.,(I) prenyltransferase complexed with a buffer molecule (TAPS), (2)prenyltransferase as a binary complex with geranyl diphosphate (GPP) andMg²⁺, and prenyltransferase as ternary complexes with a non-hydrolyzablesubstrate analogue, geranyl S-thiolodiphosphate (GSPP) and either (3)1,6-dihydroxynaphthalene (1,6-DHN), or (4) flaviolin (i.e.,2,5,7-trihydroxy-1,4-naphthoquinone, which is the oxidized product of1,3,6,8-tetrahydroxynaphthalene (THN)). These structures have beensolved and refined to 1.5 Å, 2.25 Å, 1.95 Å and 2.09 Å, respectively.This first structure of an aromatic prenyltransferase displays anunexpected and non-canonical (β/α)-barrel architecture.

The complexes with both aromatic substrates and geranyl containingsubstrates and analogs delineate the active site and are consistent witha proposed electrophilic mechanism of prenyl group transfer. Thesestructures also provide a mechanistic basis for understanding prenylchain length determination and substrate recognition in thisstructurally unique family of aromatic prenyltransferases. Thisstructural information is useful for predicting the aromaticprenyltransferase activity of proteins.

Specifically, the present disclosure describes the identification of twonovel aromatic prenyltransferases with promiscuous activity: Orf2 fromStreptomyces CL.190 and HypSc from Streptomyces coelicolor. The presentdisclosure also describes a high resolution structure of a new type ofβ/α-barrel which provides a useful structural template for understandingthe mechanistic features accompanying Orf2's promiscuous activity withrespect to a number of aromatic prenyl acceptors and its means ofregulating prenyl chain length specificity through a well ordered prenylchain binding surface. The β/α-barrel catalyzes the prenylation ofaromatic compounds, accepts a wide range of aromatic substrates and useshydrophobic interactions to bind the hydrocarbon moiety of an allylicdiphosphate substrate (GPP or FPP).

It is demonstrated herein that this “biosynthetic barrel” can be used asstarting point for engineering the prenylation of natural products ofboth microbial and plant origin. The structural details involved insubstrate specificity in this newly characterized small moleculeprenyltransferase enables the biosynthetic diversification of numerousaromatic compounds found in nature, and of synthetic origin by providinga structurally guided process of enzyme design and evolution, leading tothe production and metabolic engineering of novel prenylated naturalproducts through in vivo transgenic approaches, or ultimately, for invitro combinatorial chemistry.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1A presents the structures of hybrid terpenoid-polyketide compoundsproduced by Actinomycetes. The synthesis of naphterpin involves theprenylation of THN, flaviolin or a derived metabolite using a GPPco-substrate. THN is produced from malonyl-CoA by the action of THNsynthase encoded by orf3. THN is readily oxidized to give a hydroquinonederivative, 2,5,7-trihydroxy-1,4-naphthoquinone (flaviolin). The THNskeleton is further modified, prenylated and incorporated into hybridterpenoid-polyketide compounds such as naphterpin, furaquinocin A,napyradiomycin A and marinone.

FIG. 1B provides structures of representative hybrid isoprenoids fromactinomycetes. Isoprenoid units are appended to naphthoquinone (seenaphterpin, marinone, neomarinone, and Q525.518), phenol (seenovobiocin), phenazine (see lavanducyanin and aestivophoenin B), andnitropyrrole (see Q509.364) residues via C-, N-, and O-linkages, asappropriate.

FIG. 1C presents a structure based multiple sequence alignment. The orf2gene product from Streptomyces sp. strain CL190, Orf2, is a 33 kDasoluble, monomeric protein comprising 307 residues. PSI-BLAST searchesrevealed strong homologies between Orf2 and three other bacterialproteins: a protein from Streptomyces coelicolor A3(2) (HypSc, accessionnumber AL939130) and the previously described 4-hydroxyphenylpyruvate:dimethylallyl transferase genes, cloQ (accession number AF329398) andnovQ (accession number AF170880), from Streptomyces roseochromogenes andStreptomyces spheroides NCIMB 11891, respectively. Residues (one-letteramino acid code) are numbered according to Orf2's sequence. Dashesrepresent insertions and deletions. This alignment has been linked withthe known Orf2 secondary structure and rendered with ESPript (accessiblevia the internet on the world wide web at the URL“prodes.toulouse.inra.fr/ESPript”). The coding is as follows: grey ongrey for residues located in the active site, white on black forresidues strictly conserved, and white on grey overlay for residues bothstrictly conserved and located in the active site. Residues bounded bygrey frames represent similar residues in the aligned sequences.

FIG. 2A illustrates the Mg²⁺ dependent prenylation of 1,6-DHN. Thereaction buffer consisted of 50 mM HEPES (pH 7.5), 5 mM 1,6-DHN, and 5mM GPP in a final volume of 20 μl. The reaction was initiated by adding20 μg of Orf2 to the assay mixture. After incubation at 25° C. for 4hours, the mixture was dried, and spotted on a silica gel TLC plate. TheTLC plate was developed with a chloroform/methanol (20:1) solventmixture. 1,6-DHN and reaction products were detected at 254 nm. Thechemical analyses of the two HPLC purified products were accomplished byboth MS and ¹H NMR analyses. In lane 1 (control), Orf2 was boiled priorto adding. The reaction mixture in lane 2 contained no MgCl₂, while 5 mMMgCl₂ was added in lane 3.

FIG. 2B illustrates the promiscuous activity of Orf2. Several assayswere conducted, employing a variety of potential substrates, i.e.,1,3-DHN (1), 1,6-DHN (2), 2,7-DHN (3), 4-HPP (4) and severalisoflavonoids and polyketide derivatives, including daidzein(7,4′-dihydroxyisoflavonone, 5), formononetein (7-hydroxy,4′-methoxyisoflavonone, 6), fisetin (3,3′,4′,7-tetrahydroxyflavone, 7),genistein (5,7,4′-trihydroxyisoflavone, 8), naringenin(5,7,4′-trihydroxyflavonone, 9), flaviolin (10), olivetol (11),olivetolic acid (12), and resveratrol (3,4′,5-trihydroxystilbene, 13).The chemical structures of four reaction products (i.e., 1,6-dihydroxy2-geranyl naphthalene (14), 1,6-dihydroxy 5-geranyl naphthalene (15),6-geranyl naringenin (16) and 7-O-geranyl naringenin (17)) weredetermined by both MS and 1H NMR analyses. The reaction buffer consistedof 50 mM HEPES (pH 7.5), 5 mM MgCl₂, and 0.1 mM GPP, 0.009 mM [¹⁴C]GPP,and 0.1 mM of each substrate, in a final volume of 20 μl. The reactionwas initiated by adding 30 μg of Orf2 to the assay mixture. Afterincubation at 25° C. for 6 hours, the mixture was dried, and spotted ona silica gel TLC plate. The TLC plate was developed with achloroform/methanol (15:1) solvent mixture. Reaction products weredetected with a [¹⁴C] imaging plate (Fuji Photo Film).

FIG. 3 collectively presents a comparison of the different types ofprotein barrel topologies. Two-dimensional topology diagrams and threedimensional views of protein barrels are from top to bottom. Eachsecondary structure element (helices represented as circles (or spiralribbons) and β-strands as triangles (or flat ribbons)) maintainsdirectionality (N to C) which is either “up” (out of the plane of thediagram) or ‘down’ (into the plane of the diagram). The direction ofelements can be deduced from the connecting lines, and also from theorientation of the strands.

FIG. 3A illustrates an α/β-barrel (e.g., an human aldo-keto reductasecomplexed with NADP⁺ and glucose 6-phosphate (pdb entry 2ACQ)).

FIG. 3B illustrates a β/α-barrel (e.g., Streptomyces sp. strain CL190Orf2 aromatic prenyltransferase complexed with GSPP, DHN2 and Mg²⁺).

FIG. 3C illustrates an α+β-barrel (e.g., a dimeric ferrodoxin-like α+βsandwich fold of the ActVA-Orf6 monooxygenase (pdb entry 1LQ9) from S.coelicolor A3).

FIG. 3D illustrates a β-barrel (e.g., human fatty acid binding protein.M-FABP, complexed with one molecule of stearic acid (pdb entry 1HMT).

FIG. 3E illustrates an α-α barrel (e.g., the β-subunit of the Rattusnorvegicus protein farnesyltransferase complexed with farnesylated Ras4Bpeptide product and farnesyl diphosphate substrate bound simultaneously(pdb entry 1KZO)).

FIG. 4 collectively presents close-up views of the Orf2 active site indifferent complexes.

FIG. 4A illustrates the following complexes: the TAPS molecule, boundGPP, and GSPP with either 1,6-DHN or flaviolin.

FIG. 4B illustrates the structure of the divalent metal binding site. Arepresentative 2f_(o)-f_(c) electron density map (rendered from anormalized map at 1.0 σ level) displays octahedral coordination of theMg²⁺ ion, where two oxygen atoms, one from Asp 62, and one from thediphosphate moiety of the GSPP molecule contribute together with fourwater molecules to the octahedral coordination geometry.

FIG. 4C provides a schematic representation of the Orf2 active site. Theside chain involved in Mg²⁺, GSPP and 1,6-DHN binding is depicted withhydrogen and coordination bonds as grey dashed lines. Black dashed linesrepresent indirect hydrogen bonds via a water molecule. This close-upview, shown in an identical orientation to that in FIG. 4A, is rotatedby 180 degrees along the vertical axis compared to that depicted in FIG.4B. The half circles depict van der Waals contacts with the twosubstrates. Tentative depth queuing coding is as follows: grey forresidues in the back of the GSPP-1,6-DHN plane, black in the same plane,and thick black for residues in the front.

FIG. 5 presents a schematic and structural representation of theproposed mechanism for aromatic prenylation in the Orf2 active site.This panel depicts the binding of the aromatic substrate next to the GPPmolecule, formation of a geranyl carbocation (noted G+), rotation of theprenyl chain into a productive conformation, electrophilic attack of thecarbocation on the aromatic ring of 1,6-DHN, formation of a σ-complex,and final proton removal by a water molecule.

FIG. 6 presents active site models for Orf2 homologs. Modeling ofCloQ/NovQ and HypSc were performed using Orf2 as a structural template.Side chains presenting potentially significant variation between thedifferent active sites are displayed and labeled. Conserved residues inthe different models include Asp 110, Lys 119, Asn 173, Tyr 175, Tyr216, and Arg 228, of which only Asp 110 and Arg 228 are displayed forclarity.

FIG. 7 relates to the functional evaluation of HypSc. Thus, HypScprenyltransferase activity was assayed as described above with respectto FIG. 2A, using 1,6-DHN as a prenyl acceptor (in each of lanes 1-4),with no prenyl acceptor in lane 1, DMAPP in lanes 2 and 3, and GPP inlane 4. No Mg²⁺ was used in lane 2. The samples were incubated overnightat room temperature.

FIG. 8 is a block diagram of a computer system contemplated for use inthe practice of the present invention.

DETAILED DESCRIPTION OF THE INVENTION

Naphterpin is a bioactive natural product (hemiterpenoidal anti-oxidantagent) produced by Streptomyces sp. strain CL190 via both the mevalonate(MVA) isoprenoid biosynthetic pathway as well as a polyketidebiosynthetic pathway (see, for example, Shin-ya, et al., in TetrahedronLett. 31, 6025-6026 (1990); Shin-ya, et al., in J. Antibiot. (Tokyo) 43,444-447 (1990); and Seto, et al, in Tetrahedron Letters 37 (44):7979(1996), see also FIG. 1A). The compound is composed of atetrahydroxynaphthalene (THN) derivative and a geranyl moiety. THN isknown in the art to be biosynthesized from 5 molecules of malonylcoenzyme A (CoA) by the action of type III polyketide synthase (THNsynthase) cloned from Streptomyces griseus (see Funa, et al., in Nature400, 897-899 (1999)) and Streptomyces coelicolor (see Izumikawa et al.,in J. Ind. Microbiol. Biotechnol. 30:510-515 (2003)). Compounds withnaphthoquinone rings, including naphterpin, furaquinocin, napyradiomycinand marinone, are biosynthesized via the symmetric polyketideintermediate 1,3,6,8-tetrahydroxynaphthalene (THN; see Shin-ya, et al.,in J. Antibiot. (Tokyo) 43, 444-447 (1990)) (FIG. 1A). In Streptomycesgriseus and Streptomyces coelicolor A3(2), THN is the product of achalcone synthase-like type III polyketide synthase (PKS), known as TIINsynthase (THNS) (Austin and Noel, Nat Prod Rep 20 (1):79-110 (2003). THNreadily (or enzymatically) oxidizes forming a hydroquinone derivative,2,5,7-trihydroxy-1,4-naphthoquinone (flaviolin), part of whichsubsequently undergoes polymerization to form a variety of coloredpolymeric compounds (Funa et al., Nature 400 (6747):897-9 (1999)).

In addition to its role in pigment production, the THN skeleton isfurther modified and incorporated into naphterpin in Streptomyces sp.strain CL190 (Shin-ya et al., J. Anitibiot (Tokyo) 45 (1):124-5 (1992)).

In actinomycetes, three mevalonate gene clusters have been cloned todate, i.e., from CL190, Kitasatospora griseola (terpentecin producer)(see Hamano, et al., in Biosci. Biotechnol. Biochem. 65:1627-1635(2001)), and Actinoplanes sp. strain A40644 (BE-40644 producer) (seeKawasaki, et al., in J. Antibiot. 56:957-966 (2003)). All of theseclusters encode mevalonate kinase, diphosphomevalonate decarboxylase,phosphomevalonate kinase, isopentenyl diphosphate isomerase,3-hydroxy-3-methylglutaryl (HMG)-CoA reductase, and HMG-CoA synthase.The order of each of the genes is also the same and respectivehomologous genes have 50 to 80% amino acid identity with each other.

In contrast to the high conservation of the mevalonate pathway geneclusters, a diversity of genes is distributed in their flanking regions.For example, the geranylgeranyl diphosphate synthase, a key enzyme ofthe terpentecin biosynthesis, is encoded in the region just upstream ofthe mevalonate kinase gene, with the terpentecin biosynthetic genecluster located further upstream. In addition, farnesyl diphosphatesynthase, a key enzyme of the BE-40644 biosynthesis, is located justupstream of the mevalonate kinase gene, with the BE-40644 biosyntheticgene cluster located in the region downstream of the mevalonate pathwaygene cluster.

These facts, taken together, gave rise to the hypothesis that themevalonate pathway genes cluster, that terpenoid biosynthetic genes areusually clustered in a terpenoid-producing actinomycetes, and that themevalonate pathway gene cluster could be a good marker to clone theterpenoid biosynthetic genes from the terpenoid-producing actinomycetes.Based on this hypothesis, in order to clone a naphterpin biosyntheticgenes cluster, the flanking regions of the mevalonate pathway genescluster which was cloned from CL190 were sequenced.

To understand the biosynthetic pathway of this mixed terpene/polyketidederived natural product, the gene cluster responsible for naphterpinproduction was identified based upon proximity to genes encoding the MVApathway biosynthetic enzymes. An upstream region of the gene clustercontaining the MVA pathway genes revealed three new open reading framesor orfs designated orf1, orf2, and orf3. The comparative analysis ofthese orfs with genes encoding functionally characterized proteins issummarized in Table 2. PSI-BLAST searches revealed homologies betweenOrf2 and three other bacterial proteins: a protein from Streptomycescoelicolor A3(2) (HypSc, accession number AL939130) and the previouslydescribed 4-hydroxyphenylpyruvate:dimethylallyl transferase genes, cloQ(accession number AF329398) and novQ (accession number AF170880), fromStreptomyces roseochromogenes and Streptomyces spheroides NCIMB 11891,respectively (FIG. 1B).

To further understand the function of the genes referred to above, amutant Streptomyces sp. (strain CL190) was prepared by disrupting theOrf2 gene. This mutant exhibited no naphterpin production. The highdegree of homology between Orf2 and the functionally characterizedprenyltransferases CloQ/NovQ (Pojer et al., Proc Natl Acad Sci USA100:2316-2321 (2003)) (FIG. 1B), and the fact that Orf3 encodes a typeIII polyketide synthase with amino acid similarity to THNS, establishesthat orf2 encodes a prenyltransferase involved in geranyl group transferto THN or a THN derivative produced through the action of orf3 (andpossibly other tailoring enzymes).

When expressed in E. coli, Orf2 is as a 33 kDa soluble, monomericprotein having 307 residues. To assess enzyme activity, the purifiedrecombinant Orf2 protein was incubated with one of the following prenyl(geranyl) donors, i.e., dimethylallyl diphosphate (DMAPP, C5), geranyldiphosphate (GPP, C10) or farnesyl diphosphate (FPP, C15) along withseveral possible substrates (i.e., prenyl acceptors) possessing one ormore aromatic groups. A variety of THN analogues (e.g.,1,3-dihydroxynaphthalene (1,3-DHN), 1,6-DHN, 2,7-DHN, and flaviolin) areobserved to function as substrates for Orf2, i.e., are converted by Orf2into prenylated derivatives thereof (see FIGS. 2A and 2B). The4-hydroxyphenylpyruvate (4-HPP) substrate of CloQ/NovQ (Pojer et al,supra) was also converted by Orf7 into a prenylated derivative thereof.In contrast, the related molecules, phenylalanine or tyrosine, did notserve as substrates (see FIG. 2B). No activity was observed with DMAPP,the highest relative activity was observed with GPP, and weak activitywas observed with FPP. In summary, Orf2 recognizes a variety ofsubstrates.

Moreover, significant Mg²⁺ dependent, in vitro activity is observed withthe dihydroxy containing THN analogs (FIGS. 2A and 2B). Thus, twoprenylated products, 1,6-DHN-P1 and 1,6-DHN-P2, were readily detected bythin layer chromatography when Orf2 was incubated with 1,6-DHN and GPP(TLC, FIGS. 2A and 2B). Large scale incubations with GPP and 1,6-DHNproduced a sufficient amount of both products (in an approximate ratioof 10:1) to permit their structure elucidation by both MS and ¹H NMRanalyses: these compounds, trans-5-geranyl 1,6-DHN and trans-2-geranyl1,6-DHN, are believed to be novel natural products (see FIG. 2A).

Orf2's potential to serve as a template for the diversification of novelaromatic natural products was demonstrated by assaying the ability ofOrf2 to interact with various flavonoids, isoflavonoids and relatedcompounds (e.g., resveratrol; see FIG. 2B). While Orf1 showedprenyltransferase activity in the presence of daidzein(7,4′-dihydroxyisoflavanone), formononetin (7-hydroxy,4′-methoxyisoflavanone), genistein (5,7,4′-trihydroxyisoflavone), andresveratrol (3,4′,5-trihydroxystilbene), little or no activity wasobserved in the same test conditions with fisetin(3,3′,4′,7-tetrahydroxyflavone). In the presence of naringenin(5,7,4′-trihydroxyflavanone) and GPP, two reaction products, 6-geranylnaringenin, and 7-O-geranyl naringenin, were identified (by both MS and¹H NMR analyses; see FIG. 2B). 6-geranyl naringenin (also known asbonannione A; see Bruno, Heterocycles 23 (5): 1147-1153 (1985)), is aprenylated flavanone displaying significant antibacterial activity(Schutz, Phytochemistry 40:1273-1277 (1995)). 7-O-geranyl naringenin,which harbors a prenyl unit in the form of an ether moiety, which isonly occasionally found in isoflavones, is a novel prenylated flavonoid.

Only a trace component in hops, 6-geranyl naringcnin is formed by theisomerization (cyclization) of the more abundant hop flavonoid,2′,4′,6′,4-tetrahydroxy-3′-geranylchalcone. Interestingly, theantifungal activity of various yellow lupin constituents has beenreported, using Cladosporium herbarum as the test fungus. It was foundthat for isoflavones, the 6-prenyl and 3′-prenyl compounds were morefungitoxic than the 8-prenyl analogues and that transformation of theprenyl group to a cyclized derivative greatly reduced or eliminated thefungitoxic effects.

Orf2 was also active in the presence of both olivetol and olivetolicacid (see FIG. 2B). These compounds are intermediates in thebiosynthesis of the therapeutic plant derived polyketide-terpene naturalproduct Δ⁹-tetrahydrocannabinol (Δ⁹-THC). Δ⁹-THC is the primarypsychoactive component found in Cannabis sativa. A synthetic analoguethereof, i.e., dronabinol, is currently used to alleviatenausea/vomiting and to stimulate appetite in order to counter weightloss in cancer and AIDS patients. The primary psychoactive component incannabis, Δ ⁹-THC affects the brain mainly by activating two specificcannabinoid receptors (CB1 and CB2). These receptors also bind to‘endogenous’ cannabinoids, which are produced naturally by the humanbody. Recent studies of the cannabinoid signaling system shows itsinvolvement in an ever-increasing number of pathological conditions. Asthe geranyl prenyltransferase activity involved in Δ⁹-THC biosynthesisin Cannabis sativa has until now only been detected in cell extracts, itwas decided to test Orf2's activity in the presence of both olivetol andolivetolic acid, two supposed intermediates of Δ⁹-THC biosynthesis: Orf2's reaction products were detected on TLC with both Δ⁹-THC precursors,differing from the C. sativa endogenous enzyme for which activity wasonly observed in the presence of olivetolate molecule. These results arevery promising for the opening of new therapeutic avenues based on theability to modulate the endocannabinoid system.

Thus, in accordance with the present invention, there are providedaromatic prenyltransferases having a beta/alpha barrel structure.

As used herein, the phrase “beta/alpha barrel structure” refers to aclosed β-sheet comprising antiparallel β-strands arranged around acentral β-barrel core, itself surrounded by a ring of α-helices formingthe outer, solvent exposed surface of the beta/alpha barrel, asdescribed in greater detail herein. Thus, aromatic prenyltransferasesare seen to have the unique beta/alpha barrel secondary structure.

This protein is the first identified and structurally characterizedenzyme involved in a mixed polyketide-isoprenoid biosynthetic pathway,namely naphterpin biosynthesis. While this protein family has beenidentified and characterized from Streptomyces bacteria, numerousprenylated aromatic natural products are found in plants. For example,the therapeutically important natural product, tetrahydrocannabinol(THC) is a mixed polyketide-isoprenoid. Biosynthetic logic would dictatethat plants are likely to contain enzymes similar in structure andfunction to Orf2, but such enzymes have thus far not been identified.(Given this likelihood, Orf2/CloQ/NovQ/HypSc are believed to be thefirst identified members of a widespread and catalytically interestingfamily of enzymes.

Exemplary aromatic prenyltransferases according to the present inventionhave the amino acid sequence set forth in SEQ ID NO:2, or conservativevariations thereof, provided that the variant polypeptide retainsprenyltransferase activity. As used herein, “conservative variations”refer to the replacement of an amino acid residue by another,biologically similar residue. Examples of conservative variationsinclude the substitution of one hydrophobic residue such as isoleucine,valine, leucine or methionine for another; or the substitution of onepolar residue for another, such as the substitution of arginine forlysine, glutamic for aspartic acids, or glutamine for asparagine, andthe like. Other illustrative examples of conservative substitutionsinclude the changes of: alanine to serine; arginine to lysine;asparagine to glutamine or histidine; aspartate to glutamate; cysteineto serine; glutamine to asparagine; glutamate to aspartate; glycine toproline; histidine to asparagine or glutamine; isoleucine to leucine orvaline; leucine to valine or isoleucine; lysine to arginine, glutamine,or glutamate; methionine to leucine or isoleucine; phenylalanine totyrosine, leucine or methionine; serine to threonine; threonine toserine; tryptophan to tyrosine; tyrosine to tryptophan or phenylalanine;valine to isoleucine or leucine, and the like. The term “conservativevariation” also includes the use of a substituted amino acid in place ofan unsubstituted amino acid.

Modifications and substitutions contemplated herein are not limited toreplacement of amino acids. For a variety of purposes, such as increasedstability, solubility, or configuration concerns, one skilled in the artwill recognize the need to introduce other modifications (e.g., bydeletion, replacement, or addition). Examples of such othermodifications include incorporation of rare amino acids, dextra-aminoacids, glycosylation sites, cytosine for specific disulfide bridgeformation. The modified peptides can be chemically synthesized, or theisolated gene can be site-directed mutagenized, or a synthetic gene canbe synthesized and expressed in bacteria, yeast, baculovirus, tissueculture, and the like.

Aromatic prenyltransferases having sequence substantially identical tothe amino acid sequence set forth in SEQ ID NO:2 are also contemplatedherein. By “substantially identical” is meant a polypeptide or nucleicacid exhibiting at least 50%, preferably 60%, more preferably 70%, morepreferably 80%, more preferably 85%, more preferably 90%, and mostpreferably 95% homology to a reference amino acid or nucleic acidsequence, provided that the “substantially identical” polypeptideretains prenyltransferase activity.

Alternatively, aromatic prenyltransferases according to the presentinvention have at least 80% sequence identity with the amino acidsequence set forth in SEQ ID NO:2. Sequence homology and identity areoften measured using sequence analysis software (e.g., Sequence AnalysisSoftware Package of the Genetics Computer Group, University of WisconsinBiotechnology Center, 1710 University Avenue, Madison, Wis. 53705). Theterm “identity” in the context of two or more nucleic acids orpolypeptide sequences, refers to two or more sequences or subsequencesthat are the same or have a specified percentage of amino acid residuesor nucleotides that are the same when compared and aligned for maximumcorrespondence over a comparison window or designated region as measuredusing any number of sequence comparison algorithms or by manualalignment and visual inspection. The term “homology” in the context oftwo or more nucleic acids or polypeptide sequences, refers to two ormore sequences or subsequences that are homologous or have a specifiedpercentage of amino acid residues or nucleotides that are homologouswhen compared and aligned for maximum correspondence over a comparisonwindow or designated region as measured using any number of sequencecomparison algorithms or by manual alignment and visual inspection.Programs as mentioned above allow for substitution of an amino acid witha similar amino acid by determining a degree of homology between thesequences being compared.

For sequence comparison, typically one sequence acts as a referencesequence, to which test sequences are compared. When using a sequencecomparison algorithm, test and reference sequences are entered into acomputer, subsequence coordinates are designated, if necessary, andsequence algorithm program parameters are designated. Default programparameters can be used, or alternative parameters can be designated. Thesequence comparison algorithm then calculates the percent sequenceidentities for the test sequences relative to the reference sequence,based on the program parameters.

A “comparison window”, as used herein, includes reference to a segment(typically having from about 20 up to about 600 contiguous residues) inwhich a sequence may be compared to a reference sequence of the samenumber of contiguous residues after the two sequences are optimallyaligned. Methods of alignment of sequence for comparison are well knownin the art. Optimal alignment of sequences for comparison can beconducted, e.g., by the local homology algorithm of Smith & Waterman,Adv. Appl. Math. 2:482 (1981), by the homology alignment algorithm ofNeedleman & Wunsch, J. Mol. Biol. 48:443 (1970), by the search forsimilarity method of Person & Lipman, Proc. Natl. Acad. Sci. USA 85:2444(1988), by computerized implementations of these algorithms (GAP,BESTFIT, FASTA, and TFASTA in the Wisconsin Genetics Software Package,Genetics Computer Group, 575 Science Dr., Madison, Wis.), or by manualalignment and visual inspection. Other algorithms for determininghomology or identity include, for example, in addition to a BLASTprogram (Basic Local Alignment Search Tool at the National Center forBiological Information), ALIGN, AMAS (Analysis of Multiply AlignedSequences), AMPS (Protein Multiple Sequence Alignment), ASSET (AlignedSegment Statistical Evaluation Tool), BANDS, BESTSCOR, BIOSCAN(Biological Sequence Comparative Analysis Node), BLIMPS (BLocks IMProvedSearcher), FASTA, Intervals & Points, BMB, CLUSTAL V, CLUSTAL W,CONSENSUS, LCONSENSUS, WCONSENSUS, Smith-Waterman algorithm, DARWIN, LasVegas algorithm FNAT (Forced Nucleotide Alignment Tool), Framealign,Framesearch, DYNAMIC, FILTER, FSAP (Fristensky Sequence AnalysisPackage), GAP (Global Alignment Program), GENAL, GIBBS, GenQuest, ISSC(Sensitive Sequence Comparison), LALIGN (Local Sequence Alignment), LCP(Local Content Program), MACAW (Multiple Alignment Construction &Analysis Workbench), MAP (Multiple Alignment Program), MBLKP, MBLKN,PIMA (Pattern-Induced Multi-sequence Alignment), SAGA (SequenceAlignment by Genetic Algorithm) and WHAT-IF. Such alignment programs canalso be used to screen genome databases to identify polynucleotidesequences having substantially identical sequences. A number of genomedatabases are available, for example, a substantial portion of the humangenome is available as part of the Human Genome Sequencing Project (J.Roach, accessible on the world wide web (www) at the URL“weber.u.Washington.edu/˜roach/human_genome_progress 2.html”) (Gibbs,1995). Several databases containing genomic information annotated withsome functional information are maintained by different organization,and are accessible via the internet on the world wide web (www), forexample, at the URL “tigr.org/tdb”; “genetics.wise.edu”;“genome-www.stanford.edu/˜ball”; “hiv-web.lan1.gov”; “ncbi.nlm.nih.gov”;“ebi.ac.uk”; “Pasteur.fr/other/biology”; and “genome.wi.mit.edu”.

One example of a useful algorithm is BLAST and BLAST 2.0 algorithms,which are described in Altschul et al., Nucl. Acids Res. 25:3389-3402(1977), and Altschul et al., J. Mol. Biol. 215:403-410 (1990),respectively. Software for performing BLAST analyses is publiclyavailable through the National Center for Biotechnology Information onthe world wide web (www) at the URL “ncbi.nlm.nih.gov”. This algorithminvolves first identifying high scoring sequence pairs (HSPs) byidentifying short words of length W in the query sequence, which eithermatch or satisfy some positive-valued threshold score T when alignedwith a word of the same length in a database sequence. T is referred toas the neighborhood word score threshold (Altschul et al., supra). Theseinitial neighborhood word hits act as seeds for initiating searches tofind longer HSPs containing them. The word hits are extended in bothdirections along each sequence for as far as the cumulative alignmentscore can be increased. Cumulative scores are calculated using, fornucleotide sequences, the parameters M (reward score for a pair ofmatching residues; always >0). For amino acid sequences, a scoringmatrix is used to calculate the cumulative score. Extension of the wordhits in each direction are halted when: the cumulative alignment scorefalls off by the quantity X from its maximum achieved value; thecumulative score goes to zero or below, due to the accumulation of oneor more negative-scoring residue alignments; or the end of eithersequence is reached. The BLAST algorithm parameters W, T, and Xdetermine the sensitivity and speed of the alignment. The BLASTN program(for nucleotide sequences) uses as defaults a wordlength (W) of 11, anexpectation (E) of 10, M=5, N=−4 and a comparison of both strands. Foramino acid sequences, the BLASTP program uses as defaults a wordlengthof 3, and expectations (E) of 10, and the BLOSUM62 scoring matrix (seeHenikoff & Henikoff, Proc. Natl. Acad. Sci. USA 89:10915 (1989))alignments (B) of 50, expectation (i) of 10, M=5, N=−4, and a comparisonof both strands.

The BLAST algorithm also performs a statistical analysis of thesimilarity between two sequences (see, e.g., Karlin & Altschul, Proc.Natl. Acad. Sci. USA 90:5873 (1993)). One measure of similarity providedby BLAST algorithm is the smallest sum probability (P(N)), whichprovides an indication of the probability by which a match between twonucleotide or amino acid sequences would occur by chance. For example, anucleic acid is considered similar to a references sequence if thesmallest sum probability in a comparison of the test nucleic acid to thereference nucleic acid is less than about 0.2, more preferably less thanabout 0.01, and most preferably less than about 0.001.

In one embodiment, protein and nucleic acid sequence homologies areevaluated using the Basic Local Alignment Search Tool (“BLAST”) Inparticular, five specific BLAST programs are used to perform thefollowing task:

-   -   (1) BLASTP and BLAST3 compare an amino acid query sequence        against a protein sequence database;    -   (2) BLASTN compares a nucleotide query sequence against a        nucleotide sequence database;    -   (3) BLASTX compares the six-frame conceptual translation        products of a query nucleotide sequence (both strands) against a        protein sequence database;    -   (4) TBLASTN compares a query protein sequence against a        nucleotide sequence database translated in all six reading        frames (both strands); and    -   (5) TBLASTX compares the six-frame translations of a nucleotide        query sequence against the six-frame translations of a        nucleotide sequence database.

The BLAST programs identify homologous sequences by identifying similarsegments, which are referred to herein as “high-scoring segment pairs,”between a query amino or nucleic acid sequence and a test sequence whichis preferably obtained from a protein or nucleic acid sequence database.High-scoring segment pairs are preferably identified (i.e., aligned) bymeans of a scoring matrix, many of which are known in the art.Preferably, the scoring matrix used is the BLOSUM62 matrix (Gonnet etal., Science 256:1443-1445 (1992); Henikoff and Henikoff, Proteins17:49-61 (1993)). Less preferably, the PAM or PAM250 matrices may alsobe used (see, e.g., Schwartz and Dayhoff, eds., Matrices for DetectingDistance Relationships: Atlas of Protein Sequence and Structure,Washington: National Biomedical Research Foundation (1978)) BLASTprograms are accessible through the U.S. National Library of Medicine,e.g., accessible on the world wide web (www) at ncbi.nlm.nih.gov.

The parameters used with the above algorithms may be adapted dependingon the sequence length and degree of homology studied. In someembodiments, the parameters may be the default parameters used by thealgorithms in the absence of instructions from the user.

In accordance with another aspect of the present invention, there areprovided nucleic acids encoding any of the above-describedprenyltransferases, including all variations embraced by the degeneracyof the genetic code. Exemplary nucleic acids according to the presentinvention include nucleic acids which specifically hybridize to thenucleotide sequence set forth in SEQ ID NO:1 (or the complement thereof)under stringent hybridization conditions, wherein said nucleic acidencodes an aromatic prenyltransferase.

Hybridization methods are well known to those skilled in the art ofmolecular biology. “Specifically hybridizable” and “specificallycomplementary” are terms that indicate a sufficient degree ofcomplementarity such that stable and specific binding occurs between afirst nucleic acid and a DNA or RNA target. The first nucleic acid neednot be 100% complementary to its target sequence to be specificallyhybridizable. A first nucleic acid is specifically hybridizable whenthere is a sufficient degree of complementarity to avoid non-specificbinding of the first nucleic acid to non-target sequences underconditions where specific binding is desired. Such binding is referredto as specific hybridization.

An alternative indication that two nucleic acid molecules are closelyrelated is that the two molecules hybridize to each other. In certainembodiments, orf2 nucleic acid variants hybridize to a disclosed orf2nucleic acid sequence (or fragments thereof), for example, under lowstringency, moderate stringency, or high stringency conditions.Hybridization conditions resulting in particular degrees of stringencywill vary depending upon the nature of the hybridization method ofchoice and the composition and length of the hybridizing nucleic acidsequences. Generally, the temperature of hybridization and the ionicstrength (for example, the Na⁺ concentration) of the hybridizationbuffer will determine the stringency of hybridization, although washtimes also influence stringency. Calculations regarding hybridizationconditions required for attaining particular degrees of stringency arediscussed by Sambrook et al. (ed.), Molecular Cloning: A LaboratoryManual, 2nd ed., vol. 1-3, Cold Spring Harbor Laboratory Press, ColdSpring Harbor, N.Y., 1989, chapters 9 and 11.

The following exemplary sets of hybridization conditions are not meantto be limiting. High stringency conditions include hybridization in5×SSC at 65° C. for 16 hours, two washes in 2×SSC at room temperature(RT) for 15 minutes each and two washes in 0.5×SSC at 65° C. for 20minutes each. Moderate stringency conditions include hybridization in5×-6×SSC at 65° C.-70° C. for 16-20 hours, two washes in 2×SSC at RT for5-20 minutes each and two washes in 1×SSC at 55° C.-70° C. for 30minutes each. Low stringency conditions include hybridization in 6×SSCat RT to 55° C. for 16-20 hours and two washes in 2×-3×SSC at RT to 55°C. for 20-30 minutes each.

Alternatively, nucleic acids according to the present invention includenucleic acids having at least 80% sequence identity with the nucleotidesequence set forth in SEQ ID NO:1, wherein said nucleic acid encodes anaromatic prenyltransferase.

To investigate the structural features accompanying prenyl chain lengthdetermination, aromatic substrate selectivity and the mechanism ofprenyl group transfer, X-ray crystal structures of four Orf2substrate/substrate analogue complexes were determined, namely Orf2complexed with a TAPS buffer molecule, a binary Orf2 complex containingGPP and Mg²⁺, a ternary Orf2 complex with a non-hydrolyzable GPPanalogue (GSPP), Mg²⁺ and 1,6-DHN, and a ternary Orf2 complex with GSPP,Mg²⁺ and flaviolin (results are summarized in Table 3).

The three dimensional structure of Orf2 consists of a single domain thatforms a novel barrel type of structure (FIG. 3). This new barrel, heretermed a β/α-barrel, is a closed β-sheet comprising sufficientantiparallel β-strands to form a central β-barrel core (typically in therange of about 6 up to 12 β-strands), with the central β-barrel coresurrounded by a ring of α-helices forming the outer, solvent exposedsurface of the barrel (FIG. 3B). In the specific example when aβ/α-barrel structure comprises 10 β-strands, the secondary connectivitynearly conforms to a (ααββ)₅ classification, but is more specificallydescribed using the (ααββ)₄-(αββ)-α nomenclature, where helices 6 and 8,both involved in inter-protein contacts in the crystal lattice, displaya helical “kink”.

The most hydrophobic section of the β/α-barrel is the region residingbetween the outer surface of the cylindrical β-barrel and the belt ofsurrounding α-helices. Additionally, a number of hydrophobic residueslocated inside the barrel accommodate the prenyl tail of the GPP andGSPP molecules, while the diphosphate or the thio-diphosphate headgroups of substrate and substrate analogs, respectively, point towardthe “upper”, more polar end of the barrel where a M²⁺ ion iscoordinated. Typically, the bottom of the barrel is capped by a shortC-terminal helix. In the specific example when a β/α-barrel structurecomprises 10 β-strands, the C-terminal helix would be α₁₁.

Structurally related proteins belonging to either the TIM barrel or theβ-barrel structural families, both of which display barrel folds withconnectivity patterns that are distinctively different from theβ/α-barrel are illustrated herein (see FIG. 3). TIM barrel proteins (ie.the aldo-keto reductase family represented by pdb entry code 2ACQ)consist of a repeated β-strand-loop-α-helix-loop motif, most oftencontaining eight repeats, with the parallel β-strands forming theinterior of an open barrel, and the helices forming the outer belt ofthe complete protein (Gerlt and Raushel, Curr Opin Chem Biol 7(2):252-64 (2003)) (FIG. 3A).

β-barrel proteins including human fatty acid-binding proteins (FABP, pdbentry code 1HMT), consist of ten anti-parallel β-strands arranged as anelliptical barrel capped at the bottom by two short α-helices(Sacchettini et al., J Mol Biol 208 (2):327-39 (1989); Xu et al., J MolBiol 268 (11):7874-84 (1993) (FIG. 3D).

Another class of protein displaying an elliptical β-barrel surrounded byhelices is the dimeric ferredoxin-like α+β sandwich fold: the ActVA-Orf6monooxygenase (pdb entry code 1LQ9) from S. coelicolor A3 belongs to thelatter class, and is a small enzyme that oxidizes a relatively largethree ringed aromatic substrate at two active sites located betweenβ-sheets and α-helices (FIG. 3C).

α/β-barrels have been defined as large structures (at least 200 aminoacids), predominantly composed of alternating α-helices and β-strands,with parallel β-strands forming a “hub” surrounded by a “tire” ofα-helices (see Branden and Tooze, Introduction to protein structure.Second edn. (1999), New York: Garland), while α+β class encompassesproteins with mainly antiparallel β-sheets but with segregated α-helicaland β-sheet regions. In this regard, it is proposed that the α/β classdefinition, including protein domains exclusively composed of parallelβ-strands, connected by α-helices, should be enlarged to include Orf2'snovel architecture, the β/α-barrel. The β/α-barrel would introduce anovel β/α-barrel category comprising antiparallel β-strands connectedand surrounded by α-helices as a subcategory of the α/β-class, butdistinct from the α/β-barrel subcategory.

Interestingly, a last type of barrel, an α-α barrel domain, can be foundin the β-subunit of the heterodimeric human protein farnesyltransferase,which catalyzes the carboxyl-terminal prenylation of Ras and severalother signaling proteins (Park et al., Science 275 (5307): 1800-4 (1997)(FIG. 3D). This domain displays a very different overall fold butpresents a similar aromatic rich substrate binding pocket and activesite topology as described herein for Orf2 (Park et al., supra; Long etal., Nature 419 (6907):645-50 (2002)).

It seems, indeed, that isoprenyl diphosphate synthases, proteinprenyltransferases, and prenyltransferases (PTases), all involved in thebinding of prenyl compounds, use a similar strategy regarding the activesite environment. In most cases, prenyl chain bonding occurs within alarge hydrophobic tunnel with highly conserved residues. Structures ofthe trans-type farnesyl diphosphate synthase display two identicalsubunits associated as a homodimer, forming a four layer helix-bundle;eight of these helices are assembled in a domain similarly to the α-αdomain previously described for the protein prenyl transferase. Thestructures of the cis-type dimeric enzymes, undecaprenyl pyrophosphatesynthase (UPPS) from E. coli and M. luteus, reveal two hydrophobictunnels each surrounded by two α-helices and four β-strands. Both UPPSenzymes require Mg²⁺ for activity even though both lack the classicalprenyl diphosphate Mg²⁺ binding motif (i.e., the (N/D)DXXD motif) foundin most other trans-prenyltransferases and terpene synthases. Thestructures of terpenoid cyclases such as pentalene synthase,5-epi-aristolochene synthase and trichodiene synthase harbor the similarstructural feature referred to as “terpenoid synthase fold” with 10-12mostly anti-parallel α-helices, as also observed in isoprenylpyrophosphate synthases and protein prenyltransferases (see Liang, Eur JBiochem 269 (14):3339-54 (2002)). All of the above cited structuresdiffer greatly from the β/α-barrel fold described herein.

While a bound TAPS molecule in the first Orf2 structure tentativelyindicated the approximate location of the diphosphate binding site nearAsp 62 (FIG. 4A), structures complexed with GPP or a non-hydrolyzableanalogue, GSPP, precisely define the residues involved in recognitionand binding of the complete GPP substrate (FIGS. 4B and 4C). Lys 119,Asn 173 and Arg 228, located near the polar open end of the barrel, arehydrogen bonded to the terminal 3-phosphate of the GSPP molecule (FIG.4C). The α-phosphate linked to the geranyl chain hydrogen bonds with Tyr216 and Lys 284, and also coordinates a Mg²⁺ ion. The completecoordination geometry of this Mg²⁺ ion exhibits perfect octahedralsymmetry, with four equatorially arranged water molecules and twoaxially located oxygen atoms contributed by the side chain carboxylateof Asp 62 and an α-phosphate non-bridging oxygen of the GSPP molecule(FIGS. 4B and 4C). Despite the absence of a (N/D)DXXD motif, a secondwell conserved residue, Asp 110, proximal to Asp 62 in the tertiarystructure, indirectly coordinates the Mg²⁺ ion via one of the fourequatorially arranged water molecules. Tyr 121 resides within hydrogenbonding distance of the bridging atom (sulfur in GSPP and oxygen in GPP)linking the diphosphate moiety to the C10 geranyl chain. Finally, thehydrophobic geranyl chain of the GPP or GSPP molecules rest against theside chains of Val 49, Phe 123, Met 162, Tyr 175 and Tyr 216 (FIG. 4C).

The ternary complexes with Mg²⁺, GSPP and either 1,6-DHN or flaviolindelineate the chemical nature of the aromatic substrate binding site(FIGS. 4A, 4B and 4C). 1,6-DHN rests against the GSPP prenyl tail and issequestered between the side chains of Met 162 and Phe 213. The Gln 295and Leu 298 side chains provided by the short C-terminal helix line thewall of the substrate binding pocket with additional contacts madethrough the side chains of Phe 213, Ser 214 and Tyr 288. Flaviolin bindsin a slightly different position than 1,6-DHN with extra pairs ofhydrogen bonds formed with Ser 214, Tyr 288 and Gln 295, while thearomatic planes of both 1,6-DHN and flaviolin reside in the same activesite orientation (FIG. 4A).

While not wishing to be bound by any theory, the structures of thesubstrates and products are consistent with an electrophilic aromaticsubstitution for the alkylation. Theoretically, two catalytic mechanismscan be considered for prenylation of aromatic substrates. One invokes acarbon mediated nucleophilic attack on the C1 carbon of GPP with thediphosphate moiety serving as a leaving group stabilized by Mg²⁺coordination and the basic character of the diphosphate binding site.This Sn2-like mechanism has been described for proteinfarnesyltransferase (Park et al., supra; Long et al., supra). A secondmechanism is reminiscent of terpene synthases involved in allylicdiphosphate biosynthesis and prenyl group cyclization and invokescarbocation mediated electrophilic capture as proposed for thetrans-prenyltransferase reaction of FPP synthase (Tarshis et al.,Biochemistry 33 (36):10871-7 (1994)) and numerous terpene synthases(cyclases) of secondary metabolism (Cane, in Comprehensive NaturalsProducts Chemistry: Isoprenoids, D. E. Cane, Editor, 1998, ElsevierScience: Oxford, UK).

The distance between the C5 atom of 1,6-DIIN, which is the identifiedsite for prenylation, or the C3 atom of flaviolin, and the C1 atom ofGSPP are 4 Å and 7 Å, respectively. Notably, these distances are similarto the 7.3 Å separation recently described in human proteinfarnesyltransferase between the C1 atom of a bound farnesyl diphosphate(FPP) molecule and a Cys residue on a peptide substrate (Long, 2002supra). Even though an Sn2-like mechanism has been proposed forprenyltransferases, these distances, combined with the structures ofsubstrates and products, and the apparent requirement for aconformational change of the cleaved prenyl chain are consistent with anelectrophilic aromatic substitution mechanism for Orf2-mediatedalkylation of aromatic substrates (see FIG. 5).

A model for the overall reaction catalyzed by Orf2 with 1,6-DHN servingas the prenyl accepting group is depicted in FIG. 5. Firstly, acarbocation intermediate is proposed to result from the ionization ofthe diphosphate moiety, triggered by Mg²⁺ coordination, electrostatichydrogen bonds with Lys 119, Arg 228, Asn 173 and Lys 284, andco-substrate binding. The positively charged C1 atom of the geranylcarbocation rotates toward the target double bond located 7 Å away onthe prenyl acceptor (as previously described for human proteinfarnesyltransferase; see also Long, 2002, supra). A “tyrosine belt”including Tyr 121, Tyr 175 and Tyr 216, surrounding the 10 carbons ofGPP, and similar to the one observed in the human proteinfarnesyltransferase (Park et al., supra; Long et al., Biochemistry 37(27):9612-8 (1998), may help stabilize and position the carbocationicintermediates via cation-π interactions (Wise and Croteau, inComprehensive Naturals Products Chemistry: Isoprenoids, D. E. Cane,Editor, 1998, Elsevier Science: Oxford, UK).

This step involves the attachment of the reactive electrophile to the C5atom of the 1,6-DHN molecule to form a resonance stabilized carbocationor σ-complex (Olah and Mo, J. Am. Chem. Soc. 94:9241 (1972)) (FIG. 5).Finally, Tyr 216, which interacts with the diphosphate moiety of the GPPmolecule, is also hydrogen bonded to a conserved and well orderednetwork of water molecules linked to the diphosphate moiety and locatedjust above the co-substrate binding location. One of these watermolecules, highlighted in FIG. 5, is ideally positioned to abstract anacidic proton from the prenylated C5 atom of the cationic σ-complexallowing for the restoration of the neutral aromatic now containing acovalently tethered geranyl chain.

To confirm the enzymatic importance of certain active site residues,preliminary mutational studies of Orf2 were carried out, and residualactivities were monitored using cell extracts containing mutant enzymes.The D62S and D62N single mutants, as well as D62S/S51R and D62N/S51Kdouble mutants, displayed only residual activity in the presence of GPP(in the presence or absence of Mg²⁺), while no detectable activity wasobserved for the D62 A single mutant with either GPP or DMAPP as aprenyl donor, indicative of the importance of D62 in catalyticprocesses.

In order to decipher the prenyl diphosphate chain length selectivity,molecular determinants of aromatic substrate recognition and divalentcation dependence, homology modeling of CloQ (Streptomycesroseochromogenes, accession number AF329398), NovQ (Streptomycesspheroides NCIMB 11891, accession number AF170880) and HypSc(Streptomyces coelicolor A3(2), accession number AL939130) sequenceswere carried out using the three dimensional architecture of Orf2 as astructural template (FIG. 6). The large degree of overall sequencesimilarity between these sequences as well as the considerable degree ofactive site conservation between Orf2 and CloQ/NovQ/HypSc is indicativeof the conservation of the β/α-barrel fold for this family of aromaticprenyltransferases.

In the HypSc model, Asp 62 is replaced by an Asn residue and iscomplemented by the replacement of Ser 52 by an Arg residue. Also, asobserved in the CloQ/NovQ model, the presence of a salt bridge betweenArg 65 and Glu 278 appears from modeling to prevent the binding of aprenyl donor with an alkyl chain longer than C5 (i.e., (C10 or C15).From this modeling analysis, the deduced protein from Streptomycescoelicolor would be predicted to show a DMAPP specificity and Mg²⁺independent prenyltransferase activity.

In order to validate this model, HypSc was subcloned from genomic DNA,and over-expressed in E. coli as an octa-histidine tagged protein andpurified for Ni²⁺-chelation chromatography. The purified enzyme was thenassayed for prenyltransferase activity using DMAPP and GPP as prenyldonors. Notable prenyltransferase activity was detected when using DMAPPand 1,6-DHN as substrates in the absence of Mg²⁺, consistent with themodel based hypothesis set forth herein regarding the chain lengthselectivity and Mg²⁺ independence of invention enzymes.

In accordance with yet another aspect of the present invention, thereare provided compositions comprising an aromatic prenyltransferase asdescribed herein in crystalline form. Optionally, such compositionsfurther comprise one or more substrates for the aromaticprenyltransferase. As used herein, “substrates” refer to compoundssusceptible to the action of invention prenyltransferases, e.g., suchreactive aromatic compounds as tetrahydroxynaphthalene, analogs,homologs and metabolites thereof.

As used herein, “analogs” refer to compounds which are related to theabove-described aromatic substrates and retain a biological activitythereof, but have one or more substitutions and/or modifications thereofrelative to the parent compound, e.g., substitution of —O— for —CH₂—.

Alternatively, analogs may have relatively little primary structuresimilarity, but may still display a biological activity of a substrateas a result of similar secondary and/or tertiary structural features,electronic properties, and the like.

As used herein, “homolog” refers to compounds which are related to theabove-described aromatic substrates by the presence or absence of asimple unit, such as a methylene unit, or some multiple of such units,e.g., —(CH₂)_(x)—.

As used herein, “metabolite” refers to compounds which are related tothe above-described substrates as a form of such compound obtained in ahuman or animal body by action of the body on the administered form ofthe compound, for example a de-methylated analogue of a compound bearinga methyl group which is obtained in the body after administration of themethylated compound as a result of action by the body on the methylatedcompound.

X-ray crystallography can elucidate the three-dimensional structure ofcrystalline forms according to the invention. Typically, the firstcharacterization of crystalline forms by X-ray crystallography candetermine the unit cell shape and its orientation in the crystal. Theterm “unit cell” refers to the smallest and simplest volume element of acrystal that is completely representative of the unit of pattern of thecrystal. The dimensions of the unit cell are defined by six numbers:dimensions a, b and c and angles α, β, and γ. A crystal can be viewed asan efficiently packed array of multiple unit cells. Detaileddescriptions of crystallographic terms are provided in Hahn, THEINTERNATIONAL TABLES FOR CRYSTALLOGRAPHY, VOLUME A, 4^(th) Ed., KluwerAcademic Publishers (1996); and Shmueli, THE INTERNATIONAL TABLES FORCRYSTALLOGRAPHY, VOLUME B, 1^(st) Ed., Kluwer Academic Publishers. Theterm “space group” refers to the symmetry of a unit cell. In a spacegroup designation (e.g., P2) the capital letter indicates the latticetype and the other symbols represent symmetry operations that can becarried out on the unit cell without changing its appearance.

The term “selenomethionine substitution” refers to the method ofproducing a chemically modified form of a protein crystal. The proteinis expressed by bacteria in media that is depleted in methionine andsupplemented with selenomethionine. Selenium is thereby incorporatedinto the crystal in place of methionine sulfurs. The location(s) ofselenium is(are) determined by X-ray diffraction analysis of thecrystal. This information is used to generate the phase information usedto construct a three-dimensional structure of the protein.

“Heavy atom derivatization” refers to a method of producing a chemicallymodified form of a protein crystal. In practice, a crystal is soaked ina solution containing heavy atom salts or organometallic compounds,e.g., lead chloride, gold thiomalate, thimerosal, uranyl acetate, andthe like, which can diffuse through the crystal and bind to theprotein's surface. Locations of the bound heavy atoms can be determinedby X-ray diffraction analysis of the soaked crystal. This information isthen used to construct phase information which can then be used toconstruct three-dimensional structures of the enzyme as described inBlundel and Johnson, PROTEIN CRYSTALLOGRAPHY. Academic Press (1976),which is incorporated by reference herein.

The knowledge obtained from X-ray diffraction patterns can be used inthe determination of the three-dimensional structure of the bindingsites of other homologous polypeptides. This is achieved through the useof commercially available software known in the art that is capable ofgenerating three-dimensional graphical representations of molecules orportions thereof from a set of structure coordinates. The binding domaincan also be predicted by various computer models. Based on thestructural X-ray coordinates of the solved structure, mutations andvariants of the solved structure can also be designed.

An exemplary isolated aromatic prenyltransferase according to thepresent invention has been further characterized by the structuralcoordinates set forth in Appendix 1.

In accordance with still another aspect of the present invention, thereare provided methods of predicting the activity and/or substratespecificity of a putative aromatic prenyltransferase, the methodscomprising:

comparing a three-dimensional representation of a known aromaticprenyltransferase and a three-dimensional representation of a putativearomatic prenyltransferase, wherein differences between the tworepresentations are predictive of aromatic prenyltransferase activityand/or substrate specificity.

In accordance with yet another aspect of the present invention, thereare provided methods of screening for compounds which bind aromaticprenyltransferase(s), said methods comprising:

modeling a potential binding agent that interacts with one or moredomains of an aromatic prenyltransferase or fragment thereof, defined bya plurality of atomic coordinates of the aromatic prenyltransferase orfragment thereof; and

determining the ability of said potential binding agent to compete withsaid aromatic prenyltransferase substrate for binding to said aromaticprenyltransferase.

As used herein, “molecular replacement” refers to generating apreliminary model of a polypeptide whose structure coordinates areunknown, by orienting and positioning a molecule whose structurecoordinates are known within the unit cell of the unknown crystal so asto best account for the observed diffraction pattern of the unknowncrystal. Phases can then be calculated from this model and combined withthe observed amplitudes to give an approximate Fourier synthesis of thestructure whose coordinates are unknown. This in turn can be subject toany of the several forms of refinement to provide a final, accuratestructure of the unknown crystal (Lattman, Meth. Enzymol. 115:55-77(1985); Rossmann, M G., ed., THE MOLECULAR REPLACEMENT METHOD (1972),Int. Sci. Rev. Ser, No. 13, Gordon & Breach, New York). Using structurecoordinates of the aromatic prenyltransferase provided herein, molecularreplacement may be used to determine the structure coordinates of acrystalline mutant, homologue, or a different crystal form of anaromatic prenyltransferase.

In accordance with this invention, an aromatic prenyltransferase, or aportion thereof, may be crystallized in association or complex with anyknown or putative substrate and/or binding agent. The crystal structuresof a series of such complexes may then be solved by molecularreplacement and compared with that of a native aromaticprenyltransferase molecule. Potential sites for modification within thearomatic prenyltransferase molecule or a corresponding substrate and/orbinding agent therefor may thus be identified based on the points ofinteraction between the aromatic prenyltransferase and substrate and/orbinding agent therefor. This information provides an additional tool fordetermining the most efficient binding interactions, for example,increased hydrophobic interactions, between an aromaticprenyltransferase and a putative chemical entity or compound, evenbefore any synthesis or modifications are performed.

All of the complexes referred to above may be studied using well-knownX-ray diffraction techniques as described herein, and may be refinedversus 2-3 Å resolution X-ray data to an R value of about 0.20 or lessusing computer software, such as X-PLOR (Yale University, 1992,distributed by Molecular Simulations, Inc.). See, e.g., Blundel &Johnson, supra; Methods in Enzymology, vol. 114 and 115, H. W. Wyckoffet al., eds., Academic Press (1985). This information may thus be usedto optimize known classes of aromatic prenyltransferase substrate and/orbinding agent therefor, such as natural THN, and to design, modifyand/or synthesize novel classes of aromatic prenyltransferase substrateand/or binding agents.

The modeling or design of substrates and/or binding agents for aromaticprenyltransferases, i.e., compounds that bind to and/or modulate anaromatic prenyltransferase polypeptide according to the inventiongenerally involves consideration of two factors. First, the compound ormolecule must be capable of physically and structurally associating withan aromatic prenyltransferase molecule. Non-covalent molecularinteractions important in the association of an aromaticprenyltransferase with a putative substrate and/or binding agent includehydrogen bonding, van der Waals and hydrophobic interactions, and thelike.

Second, the compound or molecule must be able to assume a conformationthat allows it to associate with an aromatic prenyltransferase molecule.Although certain portions of the compound or molecule will not directlyparticipate in this association, those portions may still influence theoverall conformation of the molecule. This, in turn, may have asignificant impact on affinity with the receptor. Such conformationalrequirements include the overall three-dimensional structure andorientation of the compound or molecule in relation to all or a portionof the binding site, or the spacing between functional groups of acompound or molecule comprising several chemical entities that directlyinteract with an aromatic prenyltransferase.

The term “modeling” as used herein, refers to analysis of theinteraction of an aromatic prenyltransferase and a known or testcompound or molecule by utilizing a computer generated representation ofthe molecules, as opposed to physical molecules.

The potential binding of a test compound with an aromaticprenyltransferase may be analyzed prior to its actual synthesis andtesting by the use of computer modeling techniques. If the theoreticalstructure of the given compound is indicative of insufficientinteraction and association between it and an aromaticprenyltransferase, synthesis and testing of the compound may beobviated. However, if computer modeling indicates a strong interaction,the molecule may then be tested for its ability to bind to an aromaticprenyltransferase. Methods of assaying for aromatic prenyltransferaseactivity are known in the art (as identified and discussed herein).Methods for assaying the effect of a potential binding agent can beperformed in the presence of a known binding agent of an aromaticprenyltransferase. For example, the effect of the potential bindingagent can be assayed by measuring the ability of the potential bindingagent to compete with a known binding agent.

A test compound may be computationally evaluated and designed by meansof a series of steps in which chemical entities or fragments arescreened and selected for their ability to associate with the individualbinding pockets or other areas of an aromatic prenyltransferaseassociated with a substrate and/or binding agent therefore.

One skilled in the art may use one of several methods to predict amolecule capable of binding to an aromatic prenyltransferase and toscreen test compounds for their ability to associate with an aromaticprenyltransferase and more particularly with the individual active site(e.g., binding pockets and/or specific points of interaction) of anaromatic prenyltransferase polypeptide. This process may begin by visualinspection of, for example, the binding pocket of an aromaticprenyltransferase on the computer screen based on structure coordinatesobtained derived from X-ray diffraction data obtained from crystals ofan aromatic prenyltransferase, such as those provided in Appendix 1.Selected fragments or chemical entities may then be positioned in avariety of orientations, or docked, within an individual binding pocketof the aromatic prenyltransferase. Docking may be accomplished usingsoftware such as Quanta and Sybyl, followed by energy minimization andmolecular dynamics with standard molecular mechanics forcefields, suchas CHARMM and AMBER.

Specialized computer programs may also assist in the process ofselecting fragments or chemical entities at this stage. These include:

-   1. GRID (Goodford, P. J., “A Computational Procedure for Determining    Energetically Favorable Binding Sites on Biologically Important    Macromolecules”, J. Med. Chem., 28, pp. 849-857 (1985)). GRID is    available from Oxford University, Oxford, UK.-   2. MCSS (Miranker, A. and M. Karplus, “Functionality Maps of Binding    Sites: A Multiple Copy Simultaneous Search Method.” Proteins:    Structure. Function and Genetics, 11, pp. 29-34 (1991)). MCSS is    available from Molecular Simulations, Burlington, Mass.-   3. AUTODOCK (Goodsell, D. S. and A. J. Olsen, “Automated Docking of    Substrates to Proteins by Simulated Annealing”, Proteins: Structure.    Function, and Genetics, 8, pp. 195-202 (1990)). AUTODOCK is    available from Scripps Research Institute, La Jolla, Calif.-   4. DOCK (Kuntz, I. D. et al., “A Geometric Approach to    Macromolecule-Ligand Interactions”, J. Mol. Biol., 161, pp. 269-288    (1982)). DOCK is available from University of California. San    Francisco, Calif.

Once suitable chemical entities or fragments have been selected, theycan be assembled into a single compound that is a candidate substrateand/or binding agent. Assembly may be performed by visual inspection ofthe relationship of the fragments to each other on the three-dimensionalimage displayed on a computer screen in relation to the structurecoordinates of the aromatic prenyltransferase molecule as set forth inAppendix 1. This would be followed by manual model building usingsoftware such as Quanta or Sybyl.

Useful programs to aid one of skill in the art in connecting theindividual chemical entities or fragments include:

-   1. CAVEAT (Bartlett, P. A. et al, “CAVEAT: A Program to Facilitate    the Structure-Derived Design of Biologically Active Molecules”. In    “Molecular Recognition in Chemical and Biological Problems”, Special    Pub., Royal Chem. Soc., 78, pp. 182-196 (1989)). CAVEAT is available    from the University of California, Berkeley, Calif.-   2. 3D Database systems such as MACCS-3D (MDL Information Systems,    San Leandro, Calif.). This area is reviewed in Martin, Y. C., “3D    Database Searching in Drug Design”, J. Med. Chem., 35, pp. 2145-2154    (1992)).-   3. HOOK (available from Molecular Simulations, Burlington, Mass.).

In addition to the method of building or identifying a substrate and/orbinding agent in a step-wise fashion one fragment or chemical entity ata time as described above, aromatic prenyltransferase substrates and/orbinding agents may be designed as a whole or “de novo” using either anempty binding pocket or optionally including some portion(s) of a knownsubstrate(s) and/or binding agent(s). These methods include:

-   1. LUDI (Bohm, H.-J., “The Computer Program LUDI: A New Method for    the De Novo Design of Enzyme Inhibitors”, J. Comp. Aid. Molec.    Design, 6, pp. 61-78 (1992)). LUDI is available from Biosym    Technologies, San Diego, Calif.-   2. LEGEND (Nishibata, Y. and A. Itai, Tetrahedron, 47, p. 8985    (1991)). LEGEND is available from Molecular Simulations, Burlington,    Mass.-   3. LeapFrog (available from Tripos Associates, St. Louis, Mo.).

Other molecular modeling techniques may also be employed in accordancewith this invention. See, e.g., Cohen, N. C. et al., “Molecular ModelingSoftware and Methods for Medicinal Chemistry”, J. Med. Chem., 33, pp.883-894 (1990). See also, Navia, M. A. and M. A. Murcko, “The Use ofStructural Information in Drug Design”, Current Opinions in StructuralBiology, 2, pp. 202-210 (1992).

Once a test compound or binding agent has been designed or selected bythe above methods, the efficiency with which that compound may bind toan aromatic prenyltransferase may be tested and optimized bycomputational evaluation.

A compound designed or selected as a putative substrate and/or bindingagent may be further computationally optimized so that in its boundstate it would preferably lack repulsive electrostatic interaction withthe target site. Such non-complementary (e.g., electrostatic)interactions include repulsive charge-charge, dipole-dipole andcharge-dipole interactions. Specifically, the sum of all electrostaticinteractions between the binding agent and an aromatic prenyltransferasewhen the substrate and/or binding agent is bound to the aromaticprenyltransferase, preferably make a neutral or favorable contributionto the enthalpy of binding.

Specific computer software is available in the art to evaluate compounddeformation energy and electrostatic interaction. Examples of programsdesigned for such uses include: Gaussian 92, revision C (M. J. Frisch,Gaussian, Inc., Pittsburgh, Pa., 1992); AMBER, version 4.0 (P. A.Kollman, University of California at San Francisco, 1994); QUANTA/CHARMM(Molecular Simulations, Inc. Burlington, Mass. 1994); and InsightII/Discover (Biosysm Technologies Inc., San Diego, Calif., 1994). Theseprograms may be implemented, for example, using a Silicon Graphicsworkstation, IRIS 4D/35 or IBM RISC/6000 workstation model 550. Otherhardware systems and software packages will be known to those skilled inthe art of which the speed and capacity are continually modified.

Other molecular modeling techniques may also be employed in accordancewith this invention. For exemplary reviews and techniques, see, e.g.,Cohen et al., “Molecular Modeling Software and Methods for MedicinalChemistry, J. Med. Chem., 33, pp. 883-894 (1990); see also, M. A. Naviaand M. A. Murcko, “The Use of Structural Information in Drug Design”,Current Opinions in Structural Biology, 2, pp. 202-210 (1992); L. M.Balbes et al., “A Perspective of Modern Methods in Computer-Aided DrugDesign”, in Reviews in Computational Chemistry, Vol. 5, K. B. Lipkowitzand D. B. Boyd, Eds., VCH, New York, pp. 337-380 (1994); see also, W. C.Guida, “Software For Structure-Based Drug Design”, Curr. Opin. Struct.Biology, 4, pp. 777-781 (1994).]

In accordance with still another aspect of the present invention, thereare provided alternate methods of screening for compounds which bindaromatic prenyltransferase(s), said methods comprising:

-   -   defining an interaction site of an aromatic prenyltransferase        based on a plurality of atomic coordinates of said aromatic        prenyltransferase;    -   modeling a potential binding agent that fits spatially into said        interaction site;    -   contacting said potential binding agent with said aromatic        prenyltransferase in the presence of an aromatic        prenyltransferase substrate; and    -   determining the ability of said potential binding agent to        compete with said aromatic prenyltransferase substrate for        binding to said aromatic prenyltransferase.

In accordance with a further aspect of the present invention, there areprovided additional methods of screening for compounds which bindaromatic prenyltransferase(s), said methods comprising:

-   -   defining an interaction site of an aromatic prenyltransferase        based on a plurality of atomic coordinates of said aromatic        prenyltransferase;    -   modeling a potential binding agent that fits spatially into said        interaction site; and    -   determining the ability of said potential binding agent to        compete with an aromatic prenyltransferase substrate for said        interaction site by contacting said potential binding agent with        said aromatic prenyltransferase in the presence of said aromatic        prenyltransferase substrate.

In accordance with a still further aspect of the present invention,there are provided additional methods of screening for compounds whichbind aromatic prenyltransferase(s), said methods comprising:

-   -   modeling a potential binding agent that fits spatially into an        interaction site of an aromatic prenyltransferase defined by a        plurality of atomic coordinates of said aromatic        prenyltransferase;    -   contacting said potential binding agent with said aromatic        prenyltransferase in the presence of an aromatic        prenyltransferase substrate; and    -   determining the ability of said potential binding agent to        compete with said aromatic prenyltransferase substrate for        binding to said aromatic prenyltransferase.

In accordance with another aspect of the present invention, there areprovided additional methods of screening for compounds which bindaromatic prenyltransferase(s), said methods comprising:

-   -   modeling a potential binding agent that fits spatially into an        interaction site of an aromatic prenyltransferase defined by a        plurality of atomic coordinates of said aromatic        prenyltransferase; and    -   determining the ability of said potential binding agent to        compete with an aromatic prenyltransferase substrate for said        interaction site by contacting said potential binding agent with        said aromatic prenyltransferase in the presence of said aromatic        prenyltransferase substrate.

In accordance with yet another aspect of the present invention, thereare provided additional methods of screening for compounds which bindaromatic prenyltransferase(s), said methods comprising:

-   -   determining the ability of a potential binding agent to compete        with an aromatic prenyltransferase substrate for binding to an        aromatic prenyltransferase, wherein the potential binding agent        is modeled to fit spatially into an aromatic prenyltransferase        interaction site defined by a plurality of atomic coordinates.

In accordance with still another aspect of the present invention, thereare provided methods of identifying potential substrate(s) of anaromatic prenyltransferase, said methods comprising:

-   -   defining an active site of said aromatic prenyltransferase based        on a plurality of atomic coordinates of said aromatic        prenyltransferase;    -   identifying a potential substrate that fits said active site;        and    -   contacting the aromatic prenyltransferase with the potential        substrate and determining its activity thereon.

In accordance with a further aspect of the present invention, there areprovided methods of screening compounds to determine whether suchcompounds are aromatic prenyltransferase substrates, said methodscomprising:

-   -   determining the points of interaction between an aromatic        prenyltransferase and a substrate or product therefor;    -   selecting compound(s) having similar interaction with said        aromatic prenyltransferase; and    -   testing the selected compound for the ability to be converted by        said aromatic prenyltransferase.

In accordance with still another aspect of the present invention, thereare provided alternate methods of screening compounds to determinewhether such compounds are aromatic prenyltransferase substrates, saidmethods comprising:

-   -   selecting compound(s) having points of interaction with said        aromatic prenyltransferase, wherein similar points of        interaction have been determined between said aromatic        prenyltransferase and a substrate or product therefor; and    -   testing the selected compound for the ability to be converted by        said aromatic prenyltransferase.

In accordance with a still further aspect of the present invention,there are provided additional methods of screening compounds todetermine whether such compounds are aromatic prenyltransferasesubstrates, said methods comprising:

-   -   testing a compound for the ability to be converted by said        aromatic prenyltransferase,    -   wherein said compound has been selected as having points of        interaction with said aromatic prenyltransferase, and    -   wherein similar points of interaction have been determined        between said aromatic prenyltransferase and a substrate or        product therefor.

In accordance with yet another aspect of the present invention there areprovided methods for stimulating the activity of an aromaticprenyltransferase, said methods comprising contacting said aromaticprenyltransferase with an effective amount of a compound identified byany of the above-described methods.

Such compounds are typically administered as part of formulationscomprising at least one of the above-described compounds in apharmaceutically acceptable carrier therefor. Exemplary pharmaceuticallyacceptable carriers include solids, solutions, emulsions, dispersions,micelles, liposomes, and the like. Optionally, the pharmaceuticallyacceptable carrier employed herein further comprises an enteric coating.

Pharmaceutically acceptable carriers contemplated for use in thepractice of the present invention are those which render inventioncompounds amenable to oral delivery, transdermal delivery, intravenousdelivery, intramuscular delivery, topical delivery, nasal delivery, andthe like.

Thus, formulations contemplated for use in the practice of the presentinvention can be used in the form of a solid, a solution, an emulsion, adispersion, a micelle, a liposome, and the like, wherein the resultingformulation contains one or more of the compounds of the presentinvention, as an active ingredient, in admixture with an organic orinorganic carrier or excipient suitable for enterable or parenteralapplications. The active ingredient may be compounded, for example, withthe usual non-toxic, pharmaceutically acceptable carriers for tablets,pellets, capsules, suppositories, solutions, emulsions, suspensions andany other suitable for use. The carriers which can be used includeglucose, lactose, gum acacia, gelatin, manitol, starch paste, magnesiumtrisilicate, talc, corn starch, keratin, colloidal silica, potatostarch, urea, medium chain length triglycerides, dextrans, and othercarriers suitable for use in manufacturing preparations, in solid,semisolid, or liquid form. In addition auxiliary, stabilizing,thickening, and coloring agents and perfumes may be used. The activecompound(s) is (are) included in the formulation in an amount sufficientto produce the desired effect upon the process or disease condition.

Formulations contemplated for use in the practice of the presentinvention containing the active ingredient may be in a form suitable fororal use, for example, as tablets, troches, lozenges, aqueous or oilysuspensions, dispersible powders or granules, emulsions, hard or softcapsules, or syrups or elixirs. Formulations intended for oral use maybe prepared according to any method known to the art for the manufactureof pharmaceutical compositions and such formulations may contain one ormore agents selected from the group consisting of a sweetening agentsuch as sucrose, lactose, or saccharin, flavoring agents such aspeppermint, oil of wintergreen or cherry, coloring agents and preservingagents in order to provide pharmaceutically elegant and palatablepreparations. Tablets containing the active ingredient in admixture withnon-toxic pharmaceutically acceptable excipients used may be, forexample (1) inert diluents such as calcium carbonate, lactose, calciumphosphate or sodium phosphate; (2) granulating and disintegrating agentssuch corn starch, potato starch or alginic acid; (3) binding agents suchas gun tragacanth, corn starch, gelatin or acacia, and (4) lubricatingagents such as magnesium stearate, steric acid or talc. The tablets maybe uncoated or they may be coated by known techniques to delaydisintegration and absorption in the gastrointestinal tract and therebyprovide a sustained action over a longer period. For example, a timedelay material such as glyceryl monostearate or glyceryl distearate maybe employed. They may also be coated by such techniques as thosedescribed in U.S. Pat. Nos. 4,256,108; 4,160,452; and 4,265,874, to formosmotic therapeutic tablets for controlled release.

In some cases, formulations contemplated for oral use may be in the formof hard gelatin capsules wherein the active ingredient is mixed withinert solid diluent(s), for example, calcium carbonate, calciumphosphate or kaolin. They may also be in the form of soft gelatincapsules wherein the active ingredient is mixed with water or an oilmedium, for example, peanut oil, liquid paraffin, or olive oil.

Formulations contemplated for use in the practice of the presentinvention may be in the form of a sterile injectable suspension. Thissuspension may be formulated according to known methods using suitabledispersing or wetting agents and suspending agents. The sterileinjectable preparation may also be a sterile injectable solution orsuspension in a non-toxic parenterally-acceptable diluent or solvent,for example, as a solution in 1,3-butanediol. Sterile, fixed oils areconventionally employed as a solvent or suspending medium. For thispurpose any bland fixed oil may be employed including synthetic mono- ordiglycerides, fatty acids, naturally occurring vegetable oils likesesame oil, coconut oil, peanut oil, cottonseed oil, etc. or syntheticfatty vehicles like ethyl oleate or the like. Buffers, preservatives,antioxidants, and the like can be incorporated as required.

Formulations contemplated for use in the practice of the presentinvention may also be administered in the form of suppositories forrectal administration of the drug. These formulations may be prepared bymixing the drug with a suitable non-irritating excipient, such as cocoabutter, synthetic glyceride esters of polyethylene glycols, which aresolid at ordinary temperatures, but liquefy and/or dissolve in therectal cavity to release the drug. Since individual subjects may presenta wide variation in severity of symptoms and each drug has its uniquetherapeutic characteristics, the precise mode of administration anddosage employed for each subject is left to the discretion of thepractitioner.

Amounts effective for the particular therapeutic goal sought will, ofcourse, depend on the severity of the condition being treated, and theweight and general state of the subject. Various general considerationstaken into account in determining the “effective amount” are known tothose of skill in the art and are described, e.g., in Gilman et al.,eds., Goodman And Gilman's: The Pharmacological Bases of Therapeutics,8th ed., Pergamon Press, 1990; and Remington's Pharmaceutical Sciences,17th ed., Mack Publishing Co., Easton, Pa., 1990, each of which isherein incorporated by reference.

The term “effective amount” as applied to compounds contemplated for usein the practice of the present invention, means the quantity necessaryto effect the desired therapeutic result, for example, a level effectiveto treat, cure, or alleviate the symptoms of a disease state for whichthe therapeutic compound is being administered, or to establishhomeostasis. Since individual subjects may present a wide variation inseverity of symptoms and each drug or active agent has its uniquetherapeutic characteristics, the precise mode of administration, dosageemployed and treatment protocol for each subject is left to thediscretion of the practitioner.

The above-described methods for stimulating activity of aromaticprenyltransferases can be applied in many situations. For example,cancer cell resistance to chemotherapy is often mediated byoverexpression of P-glycoprotein, a plasma membrane ABC (ATP-bindingcassette) transporter which extrudes cytotoxic drugs at the expense ofATP hydrolysis. Prenylated flavonoids have recently been reported aspotential inhibitor of human multidrug resistant protein (MRP1) whichbelong to the ABC transporter superfamily. Some of these prenylatedcompounds have also shown HIV-inhibitory effects. Recently, common formsof the prenylated flavonoids have been identified in beer: 6- and8-prenylnaringenin, xanthohumol and isoxanthohumol are present in highconcentrations in hops (Humulus lupulus L.) and their oestrogenicpotency has been determined in in vitro and animal model systems, withdata indicating that they are more potent oestrogens than theisoflavones class (see Milligan et al., J Clin Endocrinol Metab84:2249-52 (1999).

In accordance with a still further aspect of the present invention,there are provided methods of identifying potential modulator(s) ofaromatic prenyltransferase(s), said methods comprising:

-   -   defining an aromatic prenyltransferase polypeptide or fragment        thereof based on a plurality of atomic coordinates of the        aromatic prenyltransferase polypeptide;    -   modeling a potential binding agent that interacts with one or        more domains of the aromatic prenyltransferase polypeptide;    -   contacting the potential binding agent with the aromatic        prenyltransferase polypeptide; and    -   determining the ability of said potential binding agent to        modulate an aromatic prenyltransferase biological function,        thereby identifying a potential modulator of an aromatic        prenyltransferase polypeptide.

As employed herein, “modulators” refers to compound(s) which, eitherdirectly (by binding to a prenyltransferase) or indirectly (as aprecursor for a compound which binds to a prenyltransferase, or aninducer which promotes production of a compound which binds to aprenyltransferase from a precursor) induce the activity ofprenyltransferase, or to repress the activity of prenyltransferase.Exemplary modulators contemplated in the practice of the presentinvention include flavonoids, isoflavonoids, and the like.

In accordance with yet another aspect of the present invention, thereare provided alternate methods of identifying potential modulator(s) ofthe activity of aromatic prenyltransferase(s), said methods comprising:

-   -   defining the active site of said aromatic prenyltransferase        based on a plurality of atomic coordinates of said aromatic        prenyltransferase,    -   contacting a potential compound that fits the active site of (a)        with the aromatic prenyltransferase in the presence of a        substrate, and    -   determining the ability of said compound to modulate the        activity of said aromatic prenyltransferase with respect to said        substrate.

In accordance with still another aspect of the present invention, thereare provided additional methods of identifying potential modulator(s) ofthe activity of aromatic prenyltransferase(s), said methods comprising:

-   -   contacting a potential compound that fits an active site based        on a plurality of atomic coordinates of said aromatic        prenyltransferase; and    -   determining the ability of said compound to modulate the        activity of said aromatic prenyltransferase.

In accordance with a further aspect of the present invention, there areprovided methods of screening for compounds that modulate the activityof aromatic prenyltransferase(s), said methods comprising:

-   -   determining the points of interaction between an aromatic        prenyltransferase, and substrate or substrate mimic therefor;    -   selecting compound(s) having similar interaction with said        aromatic prenyltransferase; and    -   testing the selected compound for the ability to modulate the        activity of an aromatic prenyltransferase.

As employed herein, “modulating” refers to the ability of a modulatorfor a prenyltransferase to either directly or indirectly induceprenyltransferase activity, or to repress prenyltransferase activity.Exemplary processes contemplated for modulation according to theinvention include cholesterol metabolism, regulation of lipidhomeostasis, stimulation of bile transport and absorption, regulation ofthe expression of genes involved in the excretion and transportation ofbile acids (including intestinal bile acid-binding protein (IBABP)),bile salt export pump (BSEP) and canalicular multi-specific organicanion transporter (cMOAT), and the like.

In accordance with still another aspect of the present invention, thereare provided alternate methods of screening for compounds that modulatethe activity of aromatic prenyltransferase(s), said methods comprising:

-   -   selecting compound(s) having points of interaction with an        aromatic prenyltransferase, wherein similar points of        interaction have been determined between said aromatic        prenyltransferase and a substrate or substrate mimic therefor;        and    -   testing the selected compound for the ability to modulate the        activity of said aromatic prenyltransferase.

In accordance with yet another aspect of the present invention, thereare provided additional methods of screening for compounds that modulatethe activity of aromatic prenyltransferase(s), said methods comprising:

-   -   testing a compound for the ability to modulate the activity of        an aromatic prenyltransferase,    -   wherein said compound has been selected as having points of        interaction with said aromatic prenyltransferase, and    -   wherein similar points of interaction have been determined        between said aromatic prenyltransferase and a substrate or        substrate mimic therefor.

In accordance with yet another aspect of the present invention, thereare provided methods for prenylating aromatic substrates, said methodscomprising:

-   -   contacting an aromatic substrate with an aromatic        prenyltransferase as described herein, under prenylating        conditions.

In accordance with still another aspect of the present invention, thereare provided methods of identifying proteins having a beta/alpha barrelstructure, said methods comprising:

-   -   comparing a three-dimensional representation of an aromatic        prenyltransferase as described herein with a three-dimensional        representation of a putative protein having a beta/alpha barrel        structure, wherein similarities between the two representations        are predictive of aromatic prenyltransferase proteins having a        beta alpha barrel structure.

In accordance with a further aspect of the present invention, there areprovided methods for controlling the degree of prenylation promoted byan aromatic prenyltransferase, said methods comprising:

-   -   altering one or more active site residues of said aromatic        prenyltransferase so as to change the dimensions of the active        site sufficiently to control the degree of prenylation promoted        by said aromatic prenyltransferase.

As used herein, “of prenylation” refers to the number of isoprenoidunits added to a substrate. This embraces prenylation at multiple sites,as well as introduction of one or more isoprenoid units at a singlesite.

In accordance with a still further aspect of the present invention,there are provided methods for modifying the degree of prenylationpromoted by an aromatic prenyltransferase, said methods comprising:

-   -   modifying one or more active site residues of said aromatic        prenyltransferase so as to change the dimensions of the active        site sufficiently to modify the degree of prenylation promoted        by said aromatic prenyltransferase.

In accordance with yet another aspect of the present invention, thereare provided methods for controlling the substrate specificity of anaromatic prenyltransferase, said methods comprising:

-   -   altering one or more active site residues of said aromatic        prenyltransferase so as to change the dimensions of the active        site sufficiently to control the selectivity of said aromatic        prenyltransferase with respect to aromatic substrates which are        prenylated by said aromatic prenyltransferase.

As used herein, “substrate specificity” refers to the selectivity withwhich an enzyme recognizes a substrate. A selective prenyltransferasewill recognize only a single, or a limited number of substrates, whereasa non-selective (promiscuous) prenyltransferase will recognize aplurality of substrates.

In accordance with a further aspect of the present invention, there areprovided methods for modifying the substrate specificity of an aromaticprenyltransferase, said methods comprising:

-   -   modifying one or more active site residues of said aromatic        prenyltransferase so as to change the dimensions of the active        site sufficiently to modify the selectivity of said aromatic        prenyltransferase with respect to aromatic substrates which are        prenylated by said aromatic prenyltransferase.

In accordance with still another aspect of the present invention, thereare provided methods for controlling the donor specificity of inaromatic prenyltransferase, said methods comprising:

-   -   altering one or more active site residues of said aromatic        prenyltransferase so as to change the dimensions of the active        site sufficiently to control the selectivity of said aromatic        prenyltransferase with respect to prenyl donors which are        employed to prenylate an aromatic substrate.

As used herein, “donor specificity” refers to the selectivity with whichan enzyme recognizes a prenyl donor. A selective prenyltransferase willrecognize only a single, or a limited number of prenyl donors, whereas anon-selective (promiscuous) prenyltransferase will recognize a pluralityof prenyl donors. Exemplary prenyl donors include dimethylallyldiphosphate (DMAPP, C5), isopentenyl diphosphate (IPP, C5), geranyldiphosphate (GPP, C10), farnesyl diphosphate (FPP, C15), and the like.

In accordance with a further aspect of the present invention, there areprovided methods for modifying the donor specificity of an aromaticprenyltransferase, said methods comprising:

-   -   modifying one or more active site residues of said aromatic        prenyltransferase so as to change the dimensions of the active        site sufficiently to modify the selectivity of said aromatic        prenyltransferase with respect to prenyl donors employed to        prenylate an aromatic substrate.

In accordance with still another aspect of the present invention, thereare provided computer programs on a computer readable medium, saidcomputer programs comprising instructions to cause a computer to definean aromatic prenyltransferase or fragment thereof based on a pluralityof atomic coordinates of the aromatic prenyltransferase.

According to another aspect of the present invention, there is provideda computer for determining at least a portion of the structurecoordinates corresponding to X-ray diffraction data obtained from anaromatic prenyl transferase molecule or molecular complex or a homologueof said aromatic prenyl transferase molecule or molecular complex, saidcomputer comprising:

-   -   (i) a computer-readable data storage medium comprising a data        storage material encoded with machine-readable data, wherein        said data comprises at least a portion of the structure        coordinates of Appendix 1;    -   (ii) a computer-readable data storage medium comprising a data        storage material encoded with computer-readable data, wherein        said data comprises X-ray diffraction data obtained from said        aromatic prenyl transferase molecule or molecular complex or a        homologue of said aromatic prenyl transferase molecule or        molecular complex;    -   (iii) a working memory for storing instructions for processing        said computer-readable data of (i) and (ii);    -   (iv) a central-processing unit coupled to said working memory        and to said computer-readable data storage medium of (i)        and (ii) for performing a Fourier transform of the machine        readable data of (i) and for processing said computer-readable        data of (ii) into structure coordinates; and    -   (v) a display coupled to said central-processing unit for        displaying said structure coordinates of said molecule or        molecular complex.

The term “computer” as used herein can be composed of a centralprocessing unit (for example, the Pentium III from Intel Corporation, orsimilar processor from Sun, Motorola, Compaq, AMD or InternationalBusiness Machines, and the like), a working memory which may berandom-access memory or core memory, mass storage memory (for example,one or more floppy disk drives, compact disk drives or magnetic tapecontaining data recorded thereon), at least one display terminal, atleast one keyboard and accompanying input and output devices andconnections therefor. The computer typically includes a mechanism forprocessing, accessing and manipulating input data. A skilled artisan canreadily appreciate that any one of the currently available computersystems are suitable. It should also be noted that the computer can belinked to other computer systems in a network or wide area network toprovide centralized access to the information contained within thecomputer.

Contemplated input devices for entering machine readable data include,for example, telephone modem lines, cable modems, CD-ROMs, a keyboard ordisk drives. The computer may advantageously include or be programmedwith appropriate software for reading the data from the data storagecomponent or input device, for example computational programs for use inrational drug design that are described in detail below. Contemplatedoutput devices include conventional systems known in the art, forexample, display terminals, printers, or disk drives for further storageof output.

Embodiments of the invention include systems (e.g., internet basedsystems), particularly computer systems which store and manipulate thecoordinate and sequence information described herein. One example of acomputer system 100 is illustrated in block diagram form in FIG. 8. Asused herein, “a computer system” refers to the hardware components,software components, and data storage components used to analyze thecoordinates and sequences such as those set forth in Appendix 1. Thecomputer system 100 typically includes a processor for processing,accessing and manipulating the sequence data. The processor 105 can beany well-known type of central processing unit, such as, for example,the Pentium III from Intel Corporation, or similar processor from othersuppliers such as Sun, Motorola, Compaq, AMD or International BusinessMachines.

Typically the computer system 100 is a general purpose system thatcomprises the processor 105 and one or more internal data storagecomponents 110 for storing data, and one or more data retrieving devicesfor retrieving the data stored oil the data storage components. Askilled artisan can readily appreciate that any one of the currentlyavailable computer systems are suitable.

In one particular embodiment, the computer system 100 includes aprocessor 105 connected to a bus which is connected to a main memory 115preferably implemented as RAM) and one or more internal data storagedevices 110, such as a hard drive and/or other computer readable mediahaving data recorded thereon. In some embodiments, the computer system100 further includes one or more data retrieving device(s) 118 forreading the data stored on the internal data storage devices 110.

The data retrieving device 118 may represent, for example, a floppy diskdrive, a compact disk drive, a magnetic tape drive, a modem capable ofconnection to a remote data storage system (e.g., via the internet), andthe like. In some embodiments, the internal data storage device 110 is aremovable computer readable medium such as a floppy disk, a compactdisk, a magnetic tape, and the like, containing control logic and/ordata recorded thereon. The computer system 100 may advantageouslyinclude or be programmed by appropriate software for reading the controllogic and/or the data from the data storage component once inserted inthe data retrieving device.

The computer system 100 includes a display 120 which is used to displayoutput to a computer user. It should also be noted that the computersystem 100 can be linked to other computer systems 125 a-c in a networkor wide area network to provide centralized access to the computersystem 100.

Software for accessing and processing the coordinate and sequences ofAppendix 1, (such as search tools, compare tools, and modeling toolsetc.) may reside in main memory 115 during execution.

Computer programs are widely available that are capable of carrying outthe activities necessary to model structures and substrates using thecrystal structure information provided herein. Examples include, but arenot limited to, the computer programs listed below:

-   -   Catalyst Databases™—an information retrieval program accessing        chemical databases such as BioByte Master File, Derwent WDI and        ACD;    -   Catalyst/HYPO™—generates models of compounds and hypotheses to        explain variations of activity with the structure of drug        candidates;    -   Ludi™—fits molecules into the active site of a protein by        identifying and matching complementary polar and hydrophobic        groups;    -   Leapfrog™—“grows” new ligands using an algorithm with parameters        under the control of the user.

In addition, various general purpose machines may be used with programswritten in accordance with the teachings herein, or it may be moreconvenient to construct more specialized apparatus to perform theoperations. However, preferably this is implemented in one or morecomputer programs executing on programmable systems each comprising atleast one processor, at least one data storage system (includingvolatile and non-volatile memory and/or storage elements), at least oneinput device, and at least one output device. The program is executed onthe processor to perform the functions described herein.

The following examples are provided to further illustrate aspects of theinvention. These examples are non-limiting and should not be construedas limiting any aspect of the invention.

EXAMPLES

All solvents and reagents were obtained from the Aldrich ChemicalCompany (Milwaukee, Wis.) unless otherwise indicated.

Example 1 Cloning of Orf2

A cosmid pCLC7 (see Takagi et al., J Bacteriol 182:41534157 (2000)),which contains the mevalonate pathway gene cluster and the flankingregions cloned from CL190, was sequenced. The DNA sequence of Orf2 wasdetermined by standard techniques, and is set forth as SEQ ID NO:1. Theamino acid sequence of Orf2 was deduced from the DNA sequence and is setforth as SEQ ID NO:2.

This sequencing revealed 3 new complete orfs, orf1, orf2, and orf3 and apartial orf4 in a 9.0 kb-BamHI-BamHI DNA fragment which containsmevalonate kinase and diphosphomevalonate decarboxylase (see pCL3301 inFIG. 6). To deduce function of each orf, a database search was done. Theresults are summarized in Table 2.

TABLE 2 Amino Most homologous proteins and their accession ORFs acidsnumbers ORF1 319 aa S. avermitilis RNA polymerase ECF-subfamily σfactor, AP005050 ORF2 307 aa S. coelicolor A3(2) protein, AL391041 ORF3410 aa S. antibioticus type III polyketide synthase, AB084489 ORF4 177aa S. erythraeus protein, AY078067

ORF2 also showed sequence similarity to the previously described4-hydroxyphenylpyruvate:dimethylallytransferase, cloQ (accession numberAF329398) and novQ (accession number AF170880), from Streptomycesroseochromogenes and Streptomyces spheroides NCIMB 11891, respectively(Pojer et al., supra). ORF3 is most likely to encode type III polyketidesynthase which produces THN. These data confirm that ORF2 encodesgeranyltransferase which catalyzes geranyl transfer to THN or a THNderivative produced by the action of ORF3.

Example 2 Cloning, Expression and Purification of the Orf2 Gene

The orf2 gene from Streptomyces sp. strain CL190 (GenBank accessionnumber AB187169) was cloned by PCR amplification of total genomic DNAfrom CL190 using oligonucleotides designed for ligation into the E. coliexpression vector pQE30 (Qiagen), to generate the expression plasmidpQEORF2. PCR amplification of pQEORF2, using oligonucleotides designedfor ligation into the E. coli expression vector pHIS8 (Jez et al.,Biochemistry 39 (5):890-902 (2000)) was carried out using the forwardprimer sequence:

5′-GGG GGG GGA TCC TCC GAA GCC GCT GAT GTC G-3′ (SEQ ID NO:3; BamHI siteunderlined), and the reverse primer sequence: 5′-GGG GGG GAA TTC TCA GTCCTC CAG CGA GTC G-3′ (SEQ ID NO:4; EcoRI site underlined)to generate the expression vector pHIS8ORF2. Constructs of pHIS8ORF2were transformed into E. coli BL21 (DE3) from Novagen. Recombinant Orf2protein was obtained and purified using a standard protocol describedbefore Jez et al., supra. Selenomethionine (Se-Met)-substituted proteinwas obtained from E. coli grown in M9 minimal medium using themethionine pathway inhibition approach (Doublié, 1997), and purified asdescribed for the native protein.

Example 3 Crystallization of Orf2

Initial crystals of the Orf2 protein (50 μm×30 μm×10 μm) were obtainedby the vapor diffusion method at 4° C. 2 μl hanging drops containing a1:1 mixture of a 15 mg.ml⁻¹ protein with crystallization buffer (28%[w/v] PEG 4000, 0.3 M magnesium nitrate, 2 mM DL-dithiothreitol (DTT),0.1 M PIPES pH 8.5) equilibrated over a 500 μl reservoir of the samesolution produced small diffracting crystals overnight. Larger crystalswere obtained by the macro-seeding technique in the same conditions.Crystals were stabilized by soaking briefly in a cryoprotectant solution(30% (w/v) PEG 4000, 15% (v/v) glycerol, 0.3 M magnesium nitrate, 2 mMDTT, 0.1 M TAPS, pH 8.5), and flash frozen in liquid nitrogen prior todata collection. Orf2 crystals belong to the P2₁2₁2 space group withaverage unit cell dimensions of a=71 Å, b=92 Å, c=48 Å, α=β=γ=90°, andcontain one monomer per asymmetric unit and a solvent content of 45%.Se-Met substituted crystals were obtained as described (Doublie, MethodsEnzymol 276:523-30 (1997)). Various complexes were obtained by soakingwild type Orf2 crystals in stabilization solution containing 5 mM GPP,10 mM GSPP and 40 mM 1,6-DHN, and 10 mM GSPP and 10 mM flaviolin (GPPand GSPP were purchased from Echelon Biosciences Inc.).

Example 4 Structure Determination and Refinement

A multi-wavelength anomalous dispersion (MAD) data set was collected atthe selenium edge on a Se-Met incorporated protein crystal at theBrookhaven National Laboratory (BNL) on beam line X8C. Data wereprocessed with HKL2000 (Otwinowski and Minor, Methods Enzymol 307-326(1997)), and reduced to a unique set of indexed intensities to aresolution of 1.6 Å. Single wavelength data sets were collected inhouse, at Brookhaven National Laboratory (BNL), the European SynchrotronFacility (ESRF), and at the Stanford Synchrotron Radiation Laboratory(SSRL) on the various complexes (Table 1). Phasing, density modificationand automatic model building were carried out with the program suiteSolve/Resolve (Terwilliger, Acta Crystallogr D Biol Crystallogr 58 (Pt11):1937-40 (2002); Terwilliger and Berendzen, Acta Crystallogr D BiolCrystallogr 55 (Pt 4):849-61 (1999)) providing a high quality initialelectron density map, using 7 identified Se sites. Additional rounds ofbuilding and refinement were carried out with the programs O (Jones, TheO Manual, 1993, Upsalla, Sweden) and CNS (Brünger, Acta Crystallogr DBiol Crystallogr 54 (Pt 5):905-21 (1998)), respectively. This firstmodel was used to solve the various complexes by molecular replacementwith AMoRe (Navaza, Acta Crystallogr D Biol Crystallogr 57 (Pt10):1367-72 (2001)).

Example 5 Detection of Prenyltransferase Activity of Orf2

The basal reaction buffer employed contained 50 mM HEPES (pH 7.5), 5 mMMgCl₂, 5 mM DTT (as needed), 5 mM prenyl acceptor (DHN1, DHN2 and DHN3;see FIG. 2B), and optionally 5 mM FPP or GPP, in a final volume of 20μl. The reaction (except for control) was initiated by adding 20 μg ofORF2 protein to the basal assay mixture. After incubation at roomtemperature for 4 hrs., the reaction mixture was dried using SpeedVacand the dried residue was spotted on a silica gel TLC plate. The platewas developed with chloroform:methanol (15-30:1). Reaction products weredetected at 254 nm UV.

With prenyl acceptors DHN1 and DHN2, and either prenyl donor, FPP orGPP, prenyltransferase activity was observed. With prenyl acceptor DHN3,prenyltransferase activity was observed with GPP.

Additional studies were carried out with ORF2 and (a) 1,6-DHN (2), (b)2,7-DHN (3), (c) daidzein (7,4′-dihydroxyisoflavonone, 5), (d) genistein(5,7,4′-trihydroxyisoflavone, 8), (e) naringenin(5,7,4′-trihydroxyflavonone, 9), (f) olivetol (12), and (g) resveratrol(3,4′,5-trihydroxystilbene, 13). These prenyl acceptors gave thefollowing reaction products:

-   -   (a) 5-geranyl-1,6-DHN and 2-geranyl-1,6-DHN;    -   (b) 1-geranyl-2,7-DHN and 1,6-digeranyl-2,7-DHN,    -   (c) 7-O-geranyl-daidzein;    -   (d) 7-O-geranyl-genistein;    -   (e) 6-geranyl-naringenin and 7-O-geranyl-naringenin;    -   (f) 2-geranyl-olivetol and 4-geranyl-olivetol, and    -   (g) 4-geranyl-resveratrol.

Example 6 Mg²⁺ Dependent Prenyltransferase Activity of Orf2

The basal reaction buffer employed contained 50 mM HEPES (pH 7.5), 5 mMDTT (as needed), 5 mM prenyl acceptor, DHN2, and 5 mM GPP, in a finalvolume of 20 μl. The magnesium-containing reaction mixture contained 5mM MgCl₂. The reaction was initiated by adding 20 μg of ORF2 protein tothe basal assay mixture. After incubation at room temperature for 4hrs., the reaction mixture was dried using SpeedVac and the driedresidue was spotted on a silica gel TLC plate. The plate was developedwith chloroform:methanol (15-30:1). Reaction products were detected at254 nm UV.

In the presence of magnesium in the reaction mixture, prenylatedproducts were readily observed, while in the absence of magnesium in thereaction mixture, no prenylated products were observed.

Example 7 Promiscuous Activity of Orf2 with Different Flavonoids andOther Compounds

The reaction buffer employed contained 50 mM HEPES (pH 7.5), 5 mM DTT(as needed), 5 mM MgCl₂, 0.1 mM of each prenyl acceptor, 0.1 mM GPP, and0.01 mM [¹⁴C]GPP in a final volume of 20 μl. The reaction was initiatedby adding 20 μg of ORF2 protein to the assay mixture. After incubationat room temperature for 4 hrs., the reaction mixture was dried usingSpeedVac and the dried residue was spotted on a silica gel TLC plate.The plate was developed with chloroform:methanol (15-30:1). Reactionproducts were detected with a phosphoimager. The compounds tested weredaidzein, fisetin, formononetin, genistein, naringenin, 4-HPP and DHN2.

With each of the prenyl acceptors tested, prenylated products werereadily observed.

Example 8 Knock Out Orf2 Mutant in Streptomyces Sp. Strain CL190

To gain insight into the function of ORF2, an ORF2 knock out mutant wasconstructed by frame-shift mutation into or orf2. Thus, the 9.0kb-BamHI-BamHI DNA fragment containing orf2 was cloned into BamHI siteof pUC118 (Takara, Kyoto, Japan), a vector of E. coli. The resultingplasmid, pCL3301, was digested with EcoRI and then a 3.5-kb EcoRI-EcoRIDNA fragment was cloned into EcoRI site of pUC118 to give pCL3301E3. Inaddition, a 2.0-kb BamHI-EcoRI DNA fragment in pCL3301 was cloned intoBamHI-EcoRI site of pBluescript (Toyobo, Tsuruga, Japan) to givepCL3301BE2. pCL3301E3 was digested with BglII, the recognition site ofwhich is in the targeted orf2, and then blunt-ended with T4 DNApolymerase (Takara). Next, this blunt-ended DNA fragment wasself-ligated to give pCL3301E3Bg, which contains orf2 having a frameshift mutation. A 3.5-kb EcoRI-EcoRI DNA fragment cut from pCL3301E3Bgwas cloned into the EcoRI site of pCL3301BE2 to give pBluedORF2.Finally, a 5.0-kb XbaI-KpnI fragment, both the recognition sites ofwhich are in the vector pBluescript, was ligated into the same sites ofpSE101 (see Dairi et al., Biosci Biotechnol Biochem 59:1835-1841(1995)), a Streptomyces-E. coli shuttle vector, to give pSEdORF2.

Streptomyces sp. strain CL190 was transformed with pSEdORF2 (asdescribed by Kieser, et al., in “Practical Streptomyces Genetics”, eds.The John Innes Foundation, Norwich (2000). General considerations aboutgene cloning in Streptomyces. pp. 211-228) and a desired transformantwas selected on R2YE plates containing 20 μg thiostrepton/ml. Next, thetransformant was cultivated in SK2 liquid medium containing 20 μgthiostrepton/ml at 30° C. for 3 days. As described by Kieser et al.,supra, protoplast was prepared from the transformant mycelium andregenerated oil R2YE medium without thiostrepton. Each regeneratedcolony was simultaneously inoculated on Bennet plates with and withoutthiostrepton and a thiostrepton sensitive colony was selected to obtainthe ORF2 knocking out mutant, Streptomyces sp. strain CL190 dORF2-8. Itwas confirmed by PCR that the mutant actually had frame-shift mutationin orf2 (FIG. 7).

The constructed mutant and CL190 were cultivated as reported by Shin-ya,et al., in J. Antibiot. (Tokyo) 43, 444-447 (1990). Myceria wereharvested by centrifuge and then naphterpin produced by CL190 wasextracted from the CL190 mycerium by the same method previously reported(Shin-ya et al., supra). The mycerium of the mutant was also extractedby the same method. Both the extracts were analyzed on silica gel-thinlayer chromatography (TLC) as described. As a result, naphterpin wasdetected in the extract from CL190, but not in the extract from themutant (FIG. 7). This result unequivocally indicates that ORF2 isessential for the naphterpin biosynthesis.

Example 9 Database Searches

Data base searches for sequence and structural homologues were performedwith PSI-BLAST and VAST (accessible via the internet on the world wideweb at the URL “ncbi.nlm.nih.gov”), SSM and DALI (accessible via theinternet on the world wide web at the URL “ebi.ac.uk/msd-srvs/ssm”), CE(accessible via the internet on the world wide web at the URL“cl.sdsc.edu/ce.html”) and DEJAVU (accessible via the internet on theworld wide web at the URL “portray.bmc.uu.se/”) respectively, throughthe Protein Data Bank (accessible via the internet on the world wide webat the URL “rcsb.org/pdb/”), the Structural Classification of Proteins(SCOP, accessible via the internet on the world wide web at the URL“scop.mrc-lmb.cam.ac.uk/scop”), and the CATH Protein structureclassification (accessible via the internet on the world wide web at theURL “biochem.ucl.ac.uk/bsm/cath”).

Example 10 Modeling

Models of CloQ/NovQ and HypSc were performed with the Modeller-4 package(Sali et al., Proteins 23 (3):318-26 (1995)) using Orf2 as a structuraltemplate (see FIG. 6). For each sequence, five different models werecalculated and evaluated. The multiple sequence alignment was then handmodified based on the superposition of the Orf2 structure with thedifferent models. Modeller was then re-run and the iteratively generatedmodels visually inspected and adjusted if necessary. Model quality wasassessed with PROCHECK (see Laskowski, et al., J Appl Cryst 26:283-291(1993)). Side chains presenting potentially significant variationbetween the different active sites are displayed and labeled. Conservedresidues in the different models include Asp 110, Lys 119, Asn 173, Tyr175, Tyr 216, and Arg 228, of which only Asp 110 and Arg 228 aredisplayed for clarity.

Example 11 Comparative Modeling of CloQ/NovQ and hypSc

While a significant degree of active site residue correspondence isconsistent with the validity of the homology models, small but criticaldifferences in key active site residues provide reasons for the shorterprenyl chain length specificity of CloQ/NovQ and for differences inaromatic substrate selectivity (Pojer et al., supra). In addition, thehomology model of HypSc is consistent with the hypothesis for prenylchain length specificity in this predicted protein. Tyr 121, involved inGPP binding in Orf2, is replaced in all other sequences by a Trp (115 inCloQ/NovQ, and 117 in HypSc): the modeled ring orientation is identicalto Tyr 121 while the increased bulkiness may better sequester theshorter C5 prenyl chain of DMAPP. Ser 64 and Gly 286, replaced by Arg(59 in CloQ/NovQ, 61 in HypSc) and Glu (274 in CloQ/NovQ/HypSC),respectively, appear poised to form an internal salt-bridge preciselyover the location of the second C5 isoprene unit of the GPP moleculeexperimentally positioned in the Orf2 active site. Identical changesobserved in the HypSc prenyltransferase model predict that this enzymewill also use DMAPP as a prenyl donor. Notably, in Orf2, the geranylchain of the GPP molecule ends next to the barrel opening, thusproviding a probable reason for Orf2's ability to accommodate the C15prenyl chain of the FPP unit.

Moreover, structural alignment of Orf2 with the CloQ/NovQ and HypScmodels reveals the molecular determinants for Orf2's requirement fordivalent cations. Asp 62, directly involved in Orf2's diphosphatebinding via a coordinated Mg²⁺ ion, is conservatively replaced in HypScby Asn 59 but changes to a Ser 57 in CloQ/NovQ. In a complementarymanner, Ser 51 in Orf2 is replaced by a positively charged Lys 47 inCloQ/NovQ and a positively charged Arg 47 in HypSc that, with littlerotamer rearrangement, can be positioned over the Mg²⁺ ion observed inOrf2. Furthermore, these basic side chains are ideally positioned forelectrostatic binding to the negatively charged α-phosphate of the GPPmolecule. However, Asp 110, involved indirectly in binding Mg²⁺ via awater molecule, is conserved in all the sequences examined (FIG. 1B);thus providing an explanation as to why CloQ and NovQ are active in theabsence of Mg²⁺ but display maximum activity in the presence of 2.5 mMMg²⁺ (Pojer et al, supra). Regarding CloQ's specificity for 4-HPP,Orf2's Gln 161 and Ser 177 are replaced by Arg, 153 and 169 in CloQ, andare positioned to possibly bind the negatively charged tail of the 4-HPPsubstrate.

This analysis allows one to predict that the HypSc enzyme is aprenyltransferase, accepting only DMAPP as a substrate, while notrequiring Mg²⁺ for its activity; this homology modeling based hypothesishas been confirmed by the cloning, protein expression and enzymaticassays of HypSc (see FIG. 7).

TABLE 3 Crystallographic data, phasing, and refinement statisticsSeMet-Orf2 Data Set λ 1 (inf. max f^(″)) λ 2 (peak, min f′) λ 3 (remote)Wt + TAPS Beam line BNL-X8C BNL-X8C BNL-X8C BNL-X6A Wavelength (Å)0.9793 0.97915 0.9641 0.934 Space Group P2₁2₁2 P2₁2₁2 P2₁2₁2 P2₁2₁2 Unitcell a, b, c (Å) 71.3, 91.2, 48.3 71.4, 91.2, 48.4 71.3, 91.1, 48.371.3, 91.2, 48.3 Resolution (Å)   50-1.55   50-1.55   50-1.50   50-1.45last shell (Å) 1.61-1.55 1.57-1.52 1.55-1.50 1.42-1.40 ObservationsOverall^(a) 156510 161052 170494 139582 Unique^(a) 86828 91336 9527449390 Redundancy^(a,b) 1.8 (1.9) 1.8 (1.5) 1.8 (1.5) 2.84 (1.94)Completeness^(a,b) (%) 98.2 (98.2) 97.5 (91.1)   98 (94.8) 78.3 (50.4)I/σ I^(b) 15.3 (2.2)  14.7 (1.7)  14.3 (1.7)  37.34 (38.9)  Rsym^(b,c)(%)  7.4 (49.9)  6.6 (48.3)  7.2 (55.7)  9.7 (51.5) No. of Se sites 7FOM^(d) centric 0.57 acentric 0.55 R_(cryst) ^(c)/R_(free) ^(f) (%)21.42 (24.22) 23.0 (25.8) Missing residues 5 6 Protein atoms 2322 2332Water molecules 254 429 Ions bound^(g) 0 0 Substrate and/or bindingagent atoms^(b) 0 15 R.m.s.d. bond length (Å) 0.005 0.005 R.m.s.d. bondangles (°) 1.2 1.2 average B-factor (Å²) protein 13.6 16.2 water 22.827.7 substrate and/or binding agent 0 20.5 Wt + GSPP + wt + GSPP + DataSet Wt + GPP 1,6-DHN Flaviolin Beam line Salk Inst. ESRF-BM30A SSRL-9.1Wavelength (Å) 1.54178 0.9797 Space Group P2₁2₁2 P2₁2₁2 P2₁2₁2 Unit cella, b, c (Å) 74.6, 91.9, 48.8 71.3, 90.2, 47.5 73.6, 91.6, 48.6Resolution (Å)   99-2.25   99-1.95   50-2.02 last shell (Å) 2.29-2.252.00-1.95 2.09-2.02 Observations Overall^(a) 70572 115225 110252Unique^(a) 16155 22960 21790 Redundancy^(a,b) 4.4 (4.2) 5.0 (5.1) 4.5(5.0) Completeness^(a,b) (%)  98 (99.9) 99.5 (99.6) 99.1 (99.5) I/σI^(b) 13.5 (2.5)  32.3 (35.5) 31.9 (38.0) Rsym^(b,c) (%)  8.6 (54.8) 8.6 (51.2)  9.0 (57.2) No. of Se sites FOM^(d) centric acentricR_(cryst) ^(c)/R_(free) ^(f) (%) 22.25 (25.1) 24.1 (27.1) 23.0 (26.8)Missing residues 5 6 6 Protein atoms 2338 2332 2332 Water molecules 205346 320 Ions bound^(g) 3 7 3 Substrate and/or binding agent atoms^(b) 1931 32 R.m.s.d. bond length (Å) 0.006 0.006 0.007 R.m.s.d. bond angles(°) 1.2 1.2 1.1 average B-factor (Å²) protein 28.2 43.5 29.4 water 35.342.5 42.6 substrate and/or binding agent 67.4 52.5 41.0 ^(a)For theSeMet data sets, F⁺ and F⁻ were considered non-equivalent whencalculating the number of unique reflections and completeness.^(b)Number in parenthesis is for highest resolution shell. ^(c)Rsym =Σ_(h) | I_(h) − <Ih> | /Σ_(h)(I_(h)), where <I_(h)> is the averageintensity over symmetry equivalent reflections. ^(d)FOM is the Figure ofMerit ^(e)R_(cryst) = Σ | | F_(obs) − F_(calc) | | /Σ | F_(obs) |, wheresummation is over the data used for refinement. ^(f)R_(free) factor isR_(cryst) calculated using 5% of data (test set) excluded fromrefinement. ^(g)Ion bounds refers to Mg²⁺, (NO₃)² ions. ^(h)Substrateand/or binding agent atoms refers to TAPS, GPP, GSPP and 1,6-DHN, GSPPand flaviolin molecules.

Thus, Table 3 summarizes the structural features accompanying prenylchain length determination, aromatic substrate selectivity and themechanism of prenyl group transfer, as determined by obtaining X-raycrystal structures of four Orf2 substrate/substrate analogue complexes,namely Orf2 complexed with a TAPS buffer molecule, a binary Orf2 complexcontaining GPP and Mg²⁺, a ternary Orf2 complex with a non-hydrolyzableGPP analogue, GSPP, Mg²⁺ and 1,6-DHN, and a ternary Orf2 complex withGSPP, Mg²⁺ and flaviolin.

Example 12 Detection of Prenyltransferase Activity of HypSc

The assay described in Example 5 was repealed with hypSc and (a) 1,6-DHN(2), (b) 2,7-DHN (3), (c) daidzein (7,4′-dihydroxyisoflavonone, 5), (d)genistein (5,7,4′-trihydroxyisoflavone, 8), (e) naringenin(5,7,4′-trihydroxyflavonone, 9), (f) olivetol (12), and (g) resveratrol(3,4′,5-trihydroxystilbene, 13). These prenyl acceptors gave thefollowing reaction products:

-   -   (a) 5-dimethylallyl-1,6-DHN;    -   (b) 1-dimethylallyl-2,7-DHN;    -   (c) no reaction products detected;    -   (d) no reaction products detected;    -   (e) 6-dimethylallyl-naringenin;    -   (f) 2-dimethylallyl-olivetol and 4-dimethylallyl-olivetol, and    -   (g) 4-dimethylallyl-resveratrol.

Example 13 Biosynthesis of Hybrid Isoprenoids from Marine Actinomycetes

With the PTases Orf2 and HypSc in hand, additional actinomycete PTaseswith different substrate specificities can be identified. To this end, agroup of marine actinomycetes that produce assorted hybrid isoprenoidnatural products was compiled (see FIG. 1B, Table 4).

TABLE 4 Hybrid isoprenoid-producing actinomycetes Strain natural productattachment isoprene arom. substrate S. sp. CL190 naphterpin C GPPhydroxynaphthalene CNB632 marinone + analogs C FPP hydroxynaphthaleneCNH099 marinone C FPP hydroxynaphthalene neomarinone C FPPhydroxynaphthalene lavanducyanin N GPP phenazine CNQ525 Q525.518 C x 2DMAPP/GPP hydroxynaphthalene CNQ509 Q509.364 O GPP phenazine Q509.366 CFPP nitropyrrole S. purpeofuscus aestivophoenins N and C DMAPP phenazine

Strain CNH099 produces three isoprenoid chemotypes, namely thefarnesylated naphterpin analog marinone, the rearranged derivativeneomarinone and the phenazine lavanducyanin. Feeding experiments withlabeled precursors delineated the biosynthetic course for thesemetabolites. The biosynthesis of the naphthoquinone core common amongstthe marinones must proceed through a symmetrical pentaketideintermediate such as THN to satisfy the observed labeling patterns.Flaviolin, a known auto-oxidation product of THN, either directly ormethylated at C10 via S-adenosyl methionine may serve as an intermediatein neomarinone biosynthesis. FPP, derived from the MEP pathway, providesthe sesquiterpenoid side chain. Prenylation may occur directly viaC-prenylation though attachment of C3 of FPP or indirectly viaO-prenylation of the C5 or C7-hydroxy groups of flaviolin followed byClaisen rearrangement to yield the same furan intermediate. Protonassisted cyclization of the linear diene following Wagner-Meerweinrearrangements yields neomarinone.

A preliminary search for the respective biosynthetic gene clustersallows the generation of a cosmid library in the E. coli-Streptomycesshuttle cosmid pOJ446, the development of a genetics system in thisstrain for homologous recombination involving the E. coli to CNH099conjugal transfer of pKC1139-based temperature-sensitive plasmids, andthe sequence analysis of genes encoding THN and phenazine biosynthesis.Additionally a pOJ446 cosmid library of the aestivophoenin producerStreptomyces purpeofuscus has been prepared. This information is usefulfor the identification and cloning of novel aromatic PTases.

Example 14 X-ray Crystallographic Structures of Orf2 Complexed toGeranylated Products

The reaction products of Orf2 incubated with GPP and 1,6-DHN andnaringenin, respectively, have been identified as trans-5-geranyl1,6-DHN/trans-2-geranyl 1,6-DHN and 6-geranyl naringenin/7-O-geranylnaringenin, respectively (see FIG. 2B). Large scale production of thesecompounds can be carried out in vitro using 500 uL reaction volumes inthe assay buffer described herein and incorporating 20-50 mM GPP and20-50 mM 1,6-DHN or (2S)-naringenin. Incubations can be carried outovernight and a sample of the resultant solution analyzed by HPLC-MS toassess the product yield and extent of reaction. Multiple reactions canbe combined (approximately 5-10 individual reactions), extracted twotimes with equal volumes of ethyl acetate each time, the combinedorganic extracts dried down, then dissolved in a minimal amount ofmethanol followed by injection on an HPLC and purified on a preparativereverse phase column. Purified products can then be characterized.Purified trans-5-geranyl 1,6-DHN, trans-2-geranyl 1,6-DHN, 6-geranylnaringenin, and 7-O-geranyl naringenin can be dissolved in 100% ethanolor methanol to near saturation (approximately 100-200 mM). Each of thefour Orf2 product complexes can be prepared employing co-crystallizationand soaking strategies. To ensure the maximal occupancy of the productin Orf2 crystals, both co-crystallization and soaking approaches employa grid whereby the concentrations of the respective products is variedbetween 5 and 25 mM.

Example 15 Creating an Orf2 Mutant Capable of Efficient Use of DMAPP andElucidate its Three Dimensional Structure in the Presence of DMASPP and1,6-DHN

In order to further define prenyl diphosphate chain length selectivity,molecular determinants of aromatic substrate recognition and divalentcation dependence, homology modeling of CloQ, NovQ and HypSc sequenceswere carried out using the three dimensional coordinates of Orf2 as astructural template (FIG. 6). While the degree of active site residuecorrespondence is consistent with the homology models discussed above,small differences in key active site residues may explain the shorterprenyl chain length specificity of CloQ/NovQ and the differences inaromatic substrate selectivity. In addition, the homology model of anewly identified PTase from Streptomyces coelicolor, HypSc, lead to thebiochemical characterization of HypSc as a DMAPP-specific,Mg²⁺-independent PTase. Prenyl chain length dependence in Orf2 can beevaluated in a variety of ways. One approach involves the generation ofa limited set of site directed mutants based upon the initial homologymodels of CloQ/NovQ and HypSc shown in FIG. 6. An alternate approachinvolves the generation of several 1024 member mutant libraries of allpossible amino acid permutations derived from the comparative analysisof Orf2 with either HypSc or CloQ/NovQ and centered around the geranylbinding site mapped previously.

The first set of Orf2 mutants can be constructed using a traditionalQuickChange protocol. Specifically, a Trp residue (residue 115 in CloQNovQ and residue 117 in HypSc) replaces Tyr 121 in Orf2. The increasedbulkiness of the indole ring in HypSc/CloQ/NovQ compared to the phenolicring in Orf2 may better accommodate the shorter C5 prenyl chain ofDMAPP. In addition, in HypSc/CloQ/NovQ, Arg and Glu residues replace Ser64 and Gly 286, respectively in Orf2 (residues 59 and 274 in CloQ/NovQand residues 61 and 274 in HypSc). This apparent salt bridge in theDMAPP-specific PTases sits poised over the location of the second C5isoprene unit of the GPP molecule. Notably, this change in the homologymodel of HypSc lead to the biochemical characterization of this newlydiscovered S. coelicolor enzyme as a DMAPP specific PTase. This ratherdirected approach towards enzyme engineering minimizes the potentialinfluence of neighboring residues towards prenyl chain lengthdetermination. If this mutagenic strategy fails to significantly alterOrf2's prenyl chain length specificity, a larger library of mutant Orf2scan be prepared employing the SCOPE approach described in Example 17.

Comparative homology modeling used to initially select residues forfurther functional examination is performed with the package Modeller-4using Orf2 as a structural template. As new sequences are identified,additional models can be constructed. For each sequence, five differentmodels are calculated and evaluated. The multiple sequence alignment isthen modified by hand based on the superposition of the Orf2 structurewith the individual models. Modeller-4 is then re-run and theiteratively generated models visually inspected and adjusted again ifnecessary. Model quality is assessed with PROCHECK.

Example 16 Development of a Quantitative PTase Kinetic Assay

To determine the steady state kinetic parameters for PTases, aradiometric TLC assay can be employed. The typical reaction buffercontemplated for use consists of 50 mM HEPES (pH 7.5), 0.1-10 mMaromatic acceptor, 0.1-5 mM [¹⁴C]-DMAPP, [¹⁴C]-GPP or [¹⁴C]-FPP (NewEngland Nuclear), 5 mM MgCl₂ in a final volume of 20 μl. The reaction isinitiated by adding 10 ng-5 μg of PTase to the assay mixture. Enzymeconcentration ranges can be selected to achieve the optimal PTaseconcentration obeying Michaelis-Menten kinetics. Incubations can becarried out and 4-6 time points collected in triplicate over an initialtime range of 1-120 minutes. Reactions can be quenched with ethylacetate. Extracts can be evaporated to dryness, re-dissolved inmethanol, and applied to Whatman LK6D silica TLC plates. The TLC platecan be developed with a chloroform/methanol (20:1) solvent mixture.Aromatic reaction products can be detected at 254 n or byautoradiography using imaging plates. Products can be quantified byscraping sections of the TLC plate into Ecolume scintillation fluid,detecting [¹⁴C]-radioactivity with a scintillation counter, andconverting the corrected cpm into nmoles of product using the finalspecific radioactivity of [¹⁴C]-DMAPP, [¹⁴C]-GPP or [¹⁴C]-FPP. Kineticconstants can be determined from initial velocity measurements, in whichproduct formation is linear over the time periods monitored (up to twohours for low activity PTases or mutants thereof). Given the fact thattwo substrates are employed, K_(M) values for the prenyl donor areestablished using saturating concentrations of aromatic acceptor(typically 50 mM) and K_(M) values for the aromatic acceptors areestablished using saturating concentrations of prenyl donors, typically50 mM. In order to reach 50 mM prenyl donor, the radioactive sample isdiluted to 50 mM using cold DMAPP, GPP or FPP and corrections fordilution applied as appropriate.

Example 17 A Rapid UPLC-MS-Based Qualitative Assay to Monitor PrenylGroup Transfer

An efficient analysis technique is desirable to serve as a qualitative(or semi-quantitative) assay for prenylation reactions. Specifically,biosynthetic transformations to be monitored include the prenylation(via IPP, GPP and FPP) of both natural and unnatural substrates forwild-type as well as mutant PTase. Efficiency for such an assay isdefined in terms of speed, resolution and sensitivity. The assay mustaccommodate large numbers of samples (high-throughput) for evaluatingthe several 1024 mutant libraries and must provide analysis on lowvolume (sub milliliter) reaction volumes (given the number of reactionsand the associated costs for reagents). In addition to screening theseenzymes and enzyme libraries against natural substrates with a selectionof isoprenoid diphosphates, screening may also be desirable with respectto various unnatural substrates designed to probe thestructure-to-reactivity relationships governing regio-specificprenylation of chemical building blocks. Finally, given the establishedpromiscuity of Orf2 and its ability to generate multiple products, theassay must also resolve and identify multiple prenylated species perreaction.

Given these requirements, Ultra Performance Liquid Chromatographycoupled with ESCi Mass Spectroscopy (UPLC-MS) has been identified as asuitable technique to satisfy the above described assay needs. Briefly,UPLC is a recent advancement in separations technology. The new 1.7 μmparticle technology coupled with operating pressures approaching 15,000psi results in gains of 1.7× in resolution, 3× in speed and 1.7× insensitivity versus standard HPLC using 5 μm particles when columnlengths are normalized. However, the greatest benefit of this technologyis realized when normalizing resolution (L/dp); here the gains are 1× inresolution, 9× in speed and 3× in sensitivity versus traditional HPLC 5μm particles. These significant gains in speed and sensitivity are verybeneficial for achieving a qualitative assay.

An additional benefit of this platform stems from the 500% reduction intime for methods development. Preferably, the experiments are carriedout using micro-well plates (96 or 384-well format) where the PTases,isoprenoid diphosphate, and aromatic substrate are sequentially addedfrom stock solutions and mixed. Libraries of mutant enzymes areconveniently purified in parallel using small scale (5-10 ml) culturesand an automated Qiagen robot for the parallel purification ofhistidine-tagged proteins. Following the optimal reaction time, thereactions are quenched and loaded into the UPLC sample organizer/managerfor assay. The target operating parameters include LC run times <5 min.(run times as short as 1 min. may be attainable) and direct injection ofthe quenched reaction mixtures to eliminate sample loss issues. Ifdirect injection is not feasible for high-throughput, an extraction stepusing a less polar organic solvent can be included, and then sample canbe taken directly from the top layer of the reaction well, even whilecovered to address any evaporation and subsequent sample concentrationissues.

Finally, product detection can be achieved via diode array UV detectiontriggering MS acquisition. Because injection volumes as low as 0.1 μlare possible, and retention times are so short, total volumes of <5ml/sample are run directly through the ESCi MS assuring all peaks aredetected. With the parent ion information, the identification ofprenylated products will be facile. The Mass Lynx data management systempermits the automation of data analysis, quickly identifying peaks ofinterest by predefining product mass tables.

Example 18 Structure Elucidation

Large scale in vitro reactions and whole cell fermentations can bedirectly analyzed on a Waters 600 HPLC or a Agilent 1100 HPLC equippedwith photodiode array detection (PDA), auto-sampling and fractioncollection. Separations are achieved using a YMC ODS-AQ 4.6×150 mmreversed-phase column with a linear solvent gradient of 0.15% TFA inwater to methanol over 30 min at a flow rate of 0.5 ml/min.Alternatively, the samples are first extracted with ethyl acetate, driedover MgSO₄, filtered, dried, and redissolved in methanol for analysis.When possible, chromatographic peaks are identified by co-injection withauthentic standards. Automated screening will be carried out on a WatersAcquity UPLC equipped with PDA detection and an in-line MicroMass ZQESCi (combination APCI-ESI) Mass Spectrometer for low resolution massanalysis. Isolation of pure constituents are carried out withpre-fractionated samples a 20×250 mm YMC pack ODS-A HPLC column that canoperate at a flow rate of up to 10 ml/min.

Structures of pure metabolites can be elucidated by 1D and 2D-NMRspectroscopy on Bruker DRX-300 and DRX-600 spectrometers, or on a VarianUnity Inova 500 Spectrometer. Proton and carbon assignments can beobtained from COSY, HSQC, HMBC, and nOe spectral data. Homonuclear ¹Hconnectivities can be determined by the phase-sensitive, double-quantumfiltered COSY experiment. One-bond heteronuclear ¹H-¹³C connectivitiescan be determined by gradient-enhanced proton-detected HSQC experiments.Two- and three-bond ¹H-¹³C connectivities can be determined bygradient-enhanced proton-detected HMBC experiments. Homonuclear ¹H nOe'scan be obtained by difference nOe experiments and by two-dimensionalROESY experiments to generate relative stereochemistry while theabsolute stereochemistry of new compounds can often be achieved throughthe modified Mosher analytical method or single crystal X-ray analysis.When appropriate, biosynthetic intermediates labeled with stableisotopes (such as sodium [1,2-¹³C₂]acetate or [U-¹³C₆]glucose) can beadministered to the cultures to aid in analog identification through the¹³C INADEQUATE or related experiment. High-resolution mass determinationcan be performed by TOF-ESI (TSRI Mass Spectroscopy Laboratory) or FAB.Additional characterization techniques include Polarimetry (Perkin-Elmer341 Polarimeter) and Fourier-Transform Infrared Spectroscopy (Nicolet4700 FT-IR).

While the invention has been described in detail with reference tocertain preferred embodiments thereof, it will be understood thatmodifications and variations are within the spirit and scope of thatwhich is described and claimed.

APPENDIX 1 PROGRAM: CNS 1 1 AUTHORS: BRUNGER, ADAMS, CLORE, DELANO,GROS, GROSSE-KUNSTLEVE, JIANG, KUSZEWSKI, NILGES, PANNU, READ, RICE,SIMONSON, WARREN DATA USED IN REFINEMENT. RESOLUTION RANGE HIGH(ANGSTROMS): 1.95 RESOLUTION RANGE LOW (ANGSTROMS): 29.74 DATA CUTOFF(SIGMA(F)): 0.0 DATA CUTOFF HIGH (ABS(F)): 1386159.12 DATA CUTOFF LOW(ABS(F)): 0.000000 COMPLETENESS(WORKING + TEST) (%): 99.4 NUMBER OFREFLECTIONS: 22923 FIT TO DATA USED IN REFINEMENT. CROSS-VALIDATIONMETHOD: THROUGHOUT FREE R VALUE TEST SET SELECTION: RANDOM R VALUE(WORKING SET): 0.241 FREE R VALUE: 0.271 FREE R VALUE TEST SET SIZE (%):5.0 FREE R VALUE TEST SET COUNT: 1154 ESTIMATED ERROR OF FREE R VALUE:0.008 FIT IN THE HIGHEST RESOLUTION BIN. TOTAL NUMBER OF BINS USED: 6BIN RESOLUTION RANGE HIGH (A): 1.95 BIN RESOLUTION RANGE LOW (A): 2.07BIN COMPLETENESS (WORKING + TEST) (%): 99.7 REFLECTIONS IN BIN (WORKINGSET): 3569 BIN R VALUE (WORKING SET): 0.352 BIN FREE R VALUE: 0.386 BINFREE R VALUE TEST SET SIZE (%): 4.9 BIN FREE R VALUE TEST SET COUNT: 184ESTIMATED ERROR OF BIN FREE R VALUE: 0.028 NUMBER OF NON-HYDROGEN ATOMSUSED IN REFINEMENT. PROTEIN ATOMS: 0 NUCLEIC ACID ATOMS: 0 HETEROGENATOMS: 0 SOLVENT ATOMS: 0 B VALUES. FROM WILSON PLOT (A**2): 25.5 MEAN BVALUE (OVERALL A**2): 43.5 OVERALL ANISOTROPIC B VALUE B11 (A**2): −3.35B22 (A**2): −13.96 B33 (A**2): 17.30 B12 (A**2): 0.00 B13 (A**2): 0.00B23 (A**2): 0.00 BULK SOLVENT MODELING. METHOD USED: FLAT MODEL KSOL:0.355809 BSOL: 42.5443 (A**2) ESTIMATED COORDINATE ERROR. ESD FROMLUZZATI PLOT (A): 0.30 ESD FROM SIGMAA (A): 0.35 LOW RESOLUTION CUTOFF(A): 5.00 CROSS VALIDATED ESTIMATED COORDINATE ERROR. ESD FROM C-VLUZZATI PLOT (A): 0.35 ESD FROM C-V SIGMAA (A): 0.36 RMS DEVIATIONS FROMIDEAL VALUES. BOND LENGTHS (A): 0.006 BOND ANGLES (DEGREES): 1.2DIHEDRAL ANGLES (DEGREES): 24.4 IMPROPER ANGLES (DEGREES): 0.89ISOTROPIC THERMAL MODEL: RESTRAINED ISOTROPIC THERMAL FACTOR RESTRAINTS.RMS SIGMA MAIN-CHAIN BOND (A**2): NULL; NULL MAIN-CHAIN ANGLE (A**2):NULL; NULL SIDE-CHAIN BOND (A**2): NULL; NULL SIDE CHAIN ANGLE (A**2):NULL; NULL NCS MODEL: NONE NCS RESTRAINTS. RMS SIGMA/WEIGHT GROUP 1POSITIONAL (A): NULL; NULL GROUP 1 B-FACTOR (A**2): NULL; NULL PARAMETERFILE 1: CNS_TOPPAR/protein_rep.param PARAMETER FILE 2:CNS_TOPPAR/dna-rna_rep.param PARAMETER FILE 3:CNS_TOPPAR/water_rep.param PARAMETER FILE 4: CNS_TOPPAR/ion.paramPARAMETER FILE 5: CNSPAR:gspp_dhn2_no3.param TOPOLOGY FILE 1:CNS_TOPPAR/protein.top TOPOLOGY FILE 2: CNS_TOPPAR/dna-rna.top TOPOLOGYFILE 3: CNS_TOPPAR/water.top TOPOLOGY FILE 4: CNS_TOPPAR/ion.topTOPOLOGY FILE 5: CNSPAR:gspp_dnh2_no3.top OTHER REFINEMENT REMARKS: NULLSEQRES  1 A 514 GLU ALA ALA ASP VAL GLU ARG VAL TYR ALA ALA MET GLUSEQRES  2 A 514 GLU ALA ALA GLY LEU LEU GLY VAL ALA CYS ALA ARG ASPSEQRES  3 A 514 LYS ILE TYR PRO LEU LEU SER THR PHE GLN ASP THR LEUSEQRES  4 A 514 VAL GLU GLY GLY SER VAL VAL VAL PHE SER MET ALA SERSEQRES  5 A 514 GLY ARG HIS SER THR GLU LEU ASP PHE SER ILE SER VALSEQRES  6 A 514 PRO THR SER HIS GLY ASP PRO TYR ALA THR VAL VAL GLUSEQRES  7 A 514 LYS GLY LEU PHE PRO ALA THR GLY HIS PRO VAL ASP ASPSEQRES  8 A 514 LEU LEU ALA ASP THR GLN LYS HIS LEU PRO VAL SER METSEQRES  9 A 514 PHE ALA ILE ASP GLY GLU VAL THR GLY GLY PHE LYS LYSSEQRES 10 A 514 THR TYR ALA PHE PHE PRO THR ASP ASN MET PRO GLY VALSEQRES 11 A 514 ALA GLU LEU SER ALA ILE PRO SER MET PRO PRO ALA VALSEQRES 12 A 514 ALA GLU ASN ALA GLU LEU PHE ALA ARG TYR GLY LEU ASPSEQRES 13 A 514 LYS VAL GLN MET THR SER MET ASP TYR LYS LYS ARG GLNSEQRES 14 A 514 VAL ASN LEU TYR PHE SER GLU LEU SER ALA GLN THR LEUSEQRES 15 A 514 GLU ALA GLU SER VAL LEU ALA LEU VAL ARG GLU LEU GLYSEQRES 16 A 514 LEU HIS VAL PRO ASN GLU LEU GLY LEU LYS PHE CYS LYSSEQRES 17 A 514 ARG SER PHE SER VAL TYR PRO THR LEU ASN TRP GLU THRSEQRES 18 A 514 GLY LYS ILE ASP ARG LEU CYS PHE ALA VAL ILE SER ASNSEQRES 19 A 514 ASP PRO THR LEU VAL PRO SER SER ASP GLU GLY ASP ILESEQRES 20 A 514 GLU LYS PHE HIS ASN TYR ALA THR LYS ALA PRO TYR ALASEQRES 21 A 514 TYR VAL GLY GLU LYS ARG THR LEU VAL TYR GLY LEU THRSEQRES 22 A 514 LEU SER PRO LYS GLU GLU TYR TYR LYS LEU GLY ALA TYRSEQRES 23 A 514 TYR HIS ILE THR ASP VAL GLN ARG GLY LEU LEU LYS ALASEQRES 24 A 514 PHE ASP MG2 GSP DH2 NO3 TIP TIP TIP TIP TIP TIP TIPSEQRES 25 A 514 TIP TIP TIP TIP TIP TIP TIP TIP TIP TIP TIP TIP TIPSEQRES 26 A 514 TIP TIP TIP TIP TIP TIP TIP TIP TIP TIP TIP TIP TIPSEQRES 27 A 514 TIP TIP TIP TIP TIP TIP TIP TIP TIP TIP TIP TIP TIPSEQRES 28 A 514 TIP TIP TIP TIP TIP TIP TIP TIP TIP TIP TIP TIP TIPSEQRES 29 A 514 TIP TIP TIP TIP TIP TIP TIP TIP TIP TIP TIP TIP TIPSEQRES 30 A 514 TIP TIP TIP TIP TIP TIP TIP TIP TIP TIP TIP TIP TIPSEQRES 31 A 514 TIP TIP TIP TIP TIP TIP TIP TIP TIP TIP TIP TIP TIPSEQRES 32 A 514 TIP TIP TIP TIP TIP TIP TIP TIP TIP TIP TIP TIP TIPSEQRES 33 A 514 TIP TIP TIP TIP TIP TIP TIP TIP TIP TIP TIP TIP TIPSEQRES 34 A 514 TIP TIP TIP TIP TIP TIP TIP TIP TIP TIP TIP TIP TIPSEQRES 35 A 514 TIP TIP TIP TIP TIP TIP TIP TIP TIP TIP TIP TIP TIPSEQRES 36 A 514 TIP TIP TIP TIP TIP TIP TIP TIP TIP TIP TIP TIP TIPSEQRES 37 A 514 TIP TIP TIP TIP TIP TIP TIP TIP TIP TIP TIP TIP TIPSEQRES 38 A 514 TIP TIP TIP TIP TIP TIP TIP TIP TIP TIP TIP TIP TIPSEQRES 39 A 514 TIP TIP TIP TIP TIP TIP TIP TIP TIP TIP TIP TIP TIPSEQRES 40 A 514 TIP TIP TIP TIP TIP TIP TIP CRYST1 71.319 90.243 47.51390.00 90.00 90.00 P 21 21 2  4 ORIGX1 1.000000 0.000000 0.000000 0.00000ORIGX2 0.000000 1.000000 0.000000 0.00000 ORIGX3 0.000000 0.0000001.000000 0.00000 SCALE1 0.014022 0.000000 0.000000 0.00000 SCALE20.000000 0.011031 0.000000 0.00000 SCALE3 0.000000 0.000000 0.0210470.00000 ATOM 1 CB GLU A 3 34.602 9.738 21.871 1.00 67.30 A ATOM 2 CG GLUA 3 34.388 9.619 23.371 1.00 67.06 A ATOM 3 CD GLU A 3 35.148 8.45323.975 1.00 66.81 A ATOM 4 OE1 GLU A 3 36.396 8.490 23.969 1.00 66.45 AATOM 5 OE2 GLU A 3 34.497 7.497 24.449 1.00 66.55 A ATOM 6 C GLU A 332.569 8.406 21.275 1.00 68.45 A ATOM 7 O GLU A 3 31.776 9.074 20.6081.00 68.75 A ATOM 8 N GLU A 3 34.407 8.713 19.628 1.00 67.62 A ATOM 9 CAGLU A 3 34.078 8.542 21.070 1.00 67.93 A ATOM 10 N ALA A 4 32.177 7.53522.198 1.00 68.54 A ATOM 11 CA ALA A 4 30.768 7.318 22.493 1.00 68.73 AATOM 12 CB ALA A 4 30.598 6.047 23.317 1.00 68.94 A ATOM 13 C ALA A 430.212 8.517 23.257 1.00 69.02 A ATOM 14 O ALA A 4 29.070 8.927 23.0451.00 69.08 A ATOM 15 N ALA A 5 31.036 9.076 24.139 1.00 68.87 A ATOM 16CA ALA A 5 30.649 10.222 24.953 1.00 67.96 A ATOM 17 CB ALA A 5 31.75210.542 25.956 1.00 67.47 A ATOM 18 C ALA A 5 30.332 11.458 24.117 1.0067.32 A ATOM 19 O ALA A 5 29.330 12.131 24.356 1.00 67.46 A ATOM 20 NASP A 6 31.186 11.762 23.144 1.00 66.37 A ATOM 21 CA ASP A 6 30.96712.927 22.291 1.00 65.49 A ATOM 22 CB ASP A 6 32.130 13.117 21.310 1.0065.78 A ATOM 23 CG ASP A 6 33.432 13.479 22.002 1.00 65.79 A ATOM 24 OD1ASP A 6 33.412 14.365 22.887 1.00 65.52 A ATOM 25 OD2 ASP A 6 34.47712.887 21.648 1.00 65.81 A ATOM 26 C ASP A 6 29.667 12.802 21.506 1.0064.46 A ATOM 27 O ASP A 6 28.836 13.710 21.519 1.00 65.11 A ATOM 28 NVAL A 7 29.493 11.675 20.824 1.00 63.25 A ATOM 29 CA VAL A 7 28.29211.442 20.031 1.00 61.69 A ATOM 30 CB VAL A 7 28.331 10.054 19.351 1.0060.95 A ATOM 31 CG1 VAL A 7 27.042 9.815 18.575 1.00 60.38 A ATOM 32 CG2VAL A 7 29.530 9.968 18.423 1.00 60.39 A ATOM 33 C VAL A 7 27.023 11.54720.871 1.00 61.64 A ATOM 34 O VAL A 7 26.055 12.185 20.458 1.00 61.60 AATOM 35 N GLU 8 27.028 10.919 22.045 1.00 61.47 A ATOM 36 CA GLU A 825.871 10.953 22.936 1.00 61.38 A ATOM 37 CB GLU A 8 26.031 9.939 24.0761.00 63.51 A ATOM 38 CG GLU A 8 25.959 8.480 23.639 1.00 67.51 A ATOM 39CD GLU A 8 26.050 7.505 24.805 1.00 70.62 A ATOM 40 OE1 GLU A 8 25.1817.560 25.706 1.00 71.88 A ATOM 41 OE2 GLU A 8 26.990 6.678 24.819 1.0072.29 A ATOM 42 C GLU A 8 25.662 12.345 23.523 1.00 59.87 A ATOM 43 OGLU A 8 24.531 12.748 23.787 1.00 59.59 A ATOM 44 N ARG A 9 26.75513.072 23.729 1.00 58.71 A ATOM 45 CA ARG A 9 26.681 14.418 24.285 1.0057.97 A ATOM 46 CB ARG A 9 28.077 14.907 24.679 1.00 57.94 A ATOM 47 CGARG A 9 28.108 16.329 25.214 1.00 58.66 A ATOM 48 CD ARG A 9 29.48116.687 25.761 1.00 59.63 A ATOM 49 NE ARG A 9 30.534 16.601 24.752 1.0060.75 A ATOM 50 CZ ARG A 9 30.662 17.435 23.724 1.00 60.72 A ATOM 51 NH1ARG A 9 29.798 18.430 23.561 1.00 59.60 A ATOM 52 NH2 ARG A 9 31.65917.277 22.861 1.00 60.10 A ATOM 53 C ARG A 9 26.054 15.381 23.281 1.0057.24 A ATOM 54 O ARG A 9 25.192 16.184 23.634 1.00 56.91 A ATOM 55 NVAL A 10 26.488 15.298 22.029 1.00 56.17 A ATOM 56 CA VAL A 10 25.94816.163 20.989 1.00 54.31 A ATOM 57 CB VAL A 10 26.769 16.052 19.688 1.0054.27 A ATOM 58 CG1 VAL A 10 26.041 16.741 18.542 1.00 54.08 A ATOM 59CG2 VAL A 10 28.130 16.691 19.891 1.00 53.91 A ATOM 60 C VAL A 10 24.48815.828 20.700 1.00 53.68 A ATOM 61 O VAL A 10 23.661 16.725 20.541 1.0053.84 A ATOM 62 N TYR A 11 24.163 14.542 20.636 1.00 52.98 A ATOM 63 CATYR A 11 22.786 14.147 20.370 1.00 52.80 A ATOM 64 CB TYR A 11 22.68312.631 21.169 1.00 52.32 A ATOM 65 CG TYR A 11 21.290 12.169 19.791 1.0051.40 A ATOM 66 CD1 TYR A 11 20.788 12.376 18.506 1.00 51.32 A ATOM 67CE1 TYR A 11 19.488 11.996 18.169 1.00 51.46 A ATOM 68 CD2 TYR A 1120.457 11.567 20.732 1.00 50.53 A ATOM 69 CE2 TYR A 11 19.158 11.18620.408 1.00 50.55 A ATOM 70 CZ TYR A 11 18.678 11.403 19.128 1.00 51.02A ATOM 71 OH TYR A 11 17.387 11.042 18.814 1.00 51.32 A ATOM 72 C TYR A11 21.882 14.570 21.528 1.00 53.23 A ATOM 73 O TYR A 11 20.705 14.86921.332 1.00 52.82 A ATOM 74 N ALA A 12 22.440 14.592 22.735 1.00 53.71 AATOM 75 CA ALA A 12 21.681 14.979 23.923 1.00 53.92 A ATOM 76 CB ALA A12 22.515 14.745 25.179 1.00 54.54 A ATOM 77 C ALA A 12 21.297 16.44723.822 1.00 53.28 A ATOM 78 O ALA A 12 20.151 16.826 24.073 1.00 53.07 AATOM 79 N ALA A 13 22.273 17.267 23.454 1.00 53.23 A ATOM 80 CA ALA A 1322.062 18.697 23.305 1.00 52.67 A ATOM 81 CB ALA A 13 23.382 19.38523.012 1.00 52.66 A ATOM 82 C ALA A 13 21.083 18.947 22.170 1.00 52.84 AATOM 83 O ALA A 13 20.266 19.865 22.239 1.00 53.13 A ATOM 84 N MET A 1421.164 18.115 21.134 1.00 52.38 A ATOM 85 CA MET A 14 20.295 18.24819.973 1.00 53.03 A ATOM 86 CB MET A 14 20.715 17.257 18.882 1.00 51.89A ATOM 87 CG MET A 14 20.349 17.708 17.474 1.00 51.33 A ATOM 88 SD MET A14 20.752 16.499 16.197 1.00 51.22 A ATOM 89 CE MET A 14 22.491 16.78615.974 1.00 49.08 A ATOM 90 C MET A 14 18.824 18.042 20.333 1.00 53.97 AATOM 91 O MET A 14 17.975 18.844 19.960 1.00 54.01 A ATOM 92 N GLU A 1518.517 16.967 21.053 1.00 55.62 A ATOM 93 CA GLU A 15 17.135 16.71021.450 1.00 57.23 A ATOM 94 CB GLU A 15 17.037 15.420 22.257 1.00 58.42A ATOM 95 CG GLU A 15 17.273 14.154 21.472 1.00 59.79 A ATOM 96 CD GLU A15 17.036 12.922 22.317 1.00 60.46 A ATOM 97 OE1 GLU A 15 17.775 12.73123.307 1.00 60.40 A ATOM 98 OE2 GLU A 15 16.106 12.152 21.995 1.00 61.48A ATOM 99 C GLU A 15 16.621 17.862 22.307 1.00 57.68 A ATOM 100 O GLU A15 15.477 18.298 22.171 1.00 57.72 A ATOM 101 N GLU A 16 17.482 18.34023.201 1.00 58.40 A ATOM 102 CA GLU A 16 17.151 19.442 24.094 1.00 59.28A ATOM 103 CB GLU A 16 18.354 19.746 24.993 1.00 60.79 A ATOM 104 CG GLUA 16 18.104 20.769 26.090 1.00 63.28 A ATOM 105 CD GLU A 16 19.29520.904 27.027 1.00 65.30 A ATOM 106 OE1 GLU A 16 19.675 19.886 27.6441.00 66.30 A ATOM 107 OE2 GLU A 16 19.853 22.019 27.143 1.00 66.14 AATOM 108 C GLU A 16 16.780 20.673 23.268 1.00 58.94 A ATOM 109 O GLU A16 15.718 21.272 23.461 1.00 58.58 A ATOM 110 N ALA A 17 17.661 21.04122.342 1.00 58.27 A ATOM 111 CA ALA A 17 17.422 22.188 21.480 1.00 56.94A ATOM 112 CB ALA A 17 18.559 22.336 20.484 1.00 57.55 A ATOM 113 C ALAA 17 16.105 21.985 20.748 1.00 56.33 A ATOM 114 O ALA A 17 15.275 22.88820.676 1.00 56.73 A ATOM 115 N ALA A 18 15.917 20.787 20.208 1.00 55.47A ATOM 116 CA ALA A 18 14.697 20.466 19.486 1.00 54.97 A ATOM 117 CB ALAA 18 14.796 19.072 18.885 1.00 54.31 A ATOM 118 C ALA A 18 13.511 20.54520.439 1.00 55.41 A ATOM 119 O ALA A 18 12.408 20.920 20.041 1.00 54.89A ATOM 120 N GLY A 19 13.749 20.186 21.698 1.00 55.81 A ATOM 121 CA GLYA 19 12.693 20.225 22.690 1.00 56.15 A ATOM 122 C GLY A 19 12.109 21.61722.812 1.00 56.43 A ATOM 123 O GLY A 19 10.896 21.800 22.726 1.00 57.40A ATOM 124 N LEU A 20 12.976 22.603 23.011 1.00 56.49 A ATOM 125 CA LEUA 20 12.538 23.985 23.139 1.00 57.06 A ATOM 126 CB LEU A 20 13.74724.925 23.087 1.00 57.40 A ATOM 127 CG LEU A 20 14.770 24.808 24.2231.00 56.87 A ATOM 128 CD1 LEU A 20 15.963 25.704 23.942 1.00 57.07 AATOM 129 CD2 LEU A 20 14.116 25.196 25.538 1.00 57.91 A ATOM 130 C LEU A20 11.546 24.355 22.038 1.00 57.57 A ATOM 131 O LEU A 20 10.574 25.06922.281 1.00 58.18 A ATOM 132 N LEU A 21 11.786 23.853 20.829 1.00 58.02A ATOM 133 CA LEU A 21 10.916 24.145 19.694 1.00 57.66 A ATOM 134 CB LEUA 21 11.729 24.134 18.399 1.00 58.07 A ATOM 135 CG LEU A 21 12.84925.173 18.313 1.00 58.73 A ATOM 136 CD1 LEU A 21 13.711 24.888 17.0971.00 59.17 A ATOM 137 CD2 LEU A 21 12.252 26.572 18.245 1.00 59.02 AATOM 138 C LEU A 21 9.749 12.172 19.565 1.00 57.77 A ATOM 139 O LEU A 218.904 23.320 18.681 1.00 57.50 A ATOM 140 N GLY A 22 9.702 22.177 20.4431.00 57.54 A ATOM 141 CA GLY A 22 8.628 21.206 20.336 1.00 56.71 A ATOM142 C GLY A 22 8.716 20.353 19.136 1.00 56.49 A ATOM 143 O GLY A 227.701 19.951 18.564 1.00 55.99 A ATOM 144 N VAL A 23 9.943 20.081 18.7071.00 56.46 A ATOM 145 CA VAL A 23 10.177 19.267 17.525 1.00 56.41 A ATOM146 CB VAL A 23 11.229 19.921 16.603 1.00 56.91 A ATOM 147 CG1 VAL A 2311.383 19.104 15.325 1.00 57.54 A ATOM 148 CG2 VAL A 23 10.819 21.35316.279 1.00 56.70 A ATOM 149 C VAL A 23 10.682 17.897 17.967 1.00 56.54A ATOM 150 O VAL A 23 11.404 17.785 18.958 1.00 56.02 A ATOM 151 N ALA A24 10.300 16.859 17.231 1.00 56.63 A ATOM 152 CA ALA A 24 10.708 15.49817.557 1.00 56.80 A ATOM 153 CB ALA A 24 9.568 14.535 17.267 1.00 55.99A ATOM 154 C ALA A 24 11.949 15.079 16.780 1.00 57.61 A ATOM 155 O ALA A24 12.082 15.386 15.593 1.00 58.66 A ATOM 156 N CYS A 25 12.857 14.38017.453 1.00 57.65 A ATOM 157 CA CYS A 25 14.081 13.911 16.817 1.00 58.14A ATOM 158 CB CYS A 25 15.257 14.002 17.783 1.00 57.35 A ATOM 159 SG CYSA 25 15.899 15.659 17.971 1.00 61.13 A ATOM 160 C CYS A 25 13.949 12.48016.323 1.00 57.86 A ATOM 161 O CYS A 25 13.127 11.714 16.824 1.00 58.16A ATOM 162 N ALA A 26 14.768 12.131 15.336 1.00 57.41 A ATOM 163 CA ALAA 26 14.765 10.793 14.759 1.00 57.03 A ATOM 164 CB ALA A 26 14.31810.854 13.301 1.00 58.04 A ATOM 165 C ALA A 26 16.166 10.208 14.854 1.0056.71 A ATOM 166 O ALA A 26 17.007 10.454 13.994 1.00 55.93 A ATOM 167 NARG A 27 16.404 9.431 15.905 1.00 57.35 A ATOM 168 CA ARG A 27 17.7038.811 16.139 1.00 57.63 A ATOM 169 CB ARG A 27 17.622 7.860 17.332 1.0058.97 A ATOM 170 CG ARG A 27 18.978 7.432 17.852 1.00 60.53 A ATOM 171CD ARG A 27 19.022 7.413 19.371 1.00 61.60 A ATOM 172 NE ARG A 27 20.3877.220 19.851 1.00 62.55 A ATOM 173 CZ ARG A 27 21.098 6.119 19.629 1.0062.39 A ATOM 174 NH1 ARG A 27 20.571 5.121 18.934 1.00 63.03 A ATOM 175NH2 ARG A 27 22.330 6.013 20.106 1.00 63.44 A ATOM 176 C ARG A 27 18.2158.045 14.926 1.00 57.62 A ATOM 177 O ARG A 27 19.418 8.009 14.667 1.0057.59 A ATOM 178 N ASP A 28 17.293 7.427 14.193 1.00 57.67 A ATOM 179 CAASP A 28 17.634 6.653 13.004 1.00 58.11 A ATOM 180 CB ASP A 28 16.3696.065 12.381 1.00 60.19 A ATOM 181 CG ASP A 28 15.645 5.131 13.316 1.0062.69 A ATOM 182 OD1 ASP A 28 16.187 4.040 13.602 1.00 65.01 A ATOM 183OD2 ASP A 28 14.535 5.492 13.769 1.00 63.34 A ATOM 184 C ASP A 28 18.3427.495 11.953 1.00 57.11 A ATOM 185 O ASP A 28 19.248 7.019 11.269 1.0056.40 A ATOM 186 N LYS A 29 17.922 8.750 11.830 1.00 56.47 A ATOM 187 CALYS A 29 18.494 9.654 10.843 1.00 55.04 A ATOM 188 CB LYS A 29 17.37510.451 10.172 1.00 55.10 A ATOM 189 CG LYS A 29 16.244 9.581 9.662 1.0056.13 A ATOM 190 CD LYS A 29 15.225 10.386 8.886 1.00 57.94 A ATOM 191CE LYS A 29 14.082 9.498 8.434 1.00 58.81 A ATOM 192 NZ LYS A 29 14.5818.276 7.733 1.00 60.48 A ATOM 193 C LYS A 29 19.531 10.614 11.411 1.0054.01 A ATOM 194 O LYS A 29 20.078 11.439 10.684 1.00 54.19 A ATOM 195 NILE A 30 19.809 10.507 12.704 1.00 52.40 A ATOM 196 CA ILE A 30 20.78211.392 13.323 1.00 51.39 A ATOM 197 CB ILE A 30 20.170 12.124 14.5331.00 51.07 A ATOM 198 CG2 ILE A 30 21.168 13.122 15.096 1.00 51.54 AATOM 199 CG1 ILE A 30 18.887 12.841 14.115 1.00 49.68 A ATOM 200 CD1 ILEA 30 19.077 13.829 12.993 1.00 49.27 A ATOM 201 C ILE A 30 22.048 10.67613.781 1.00 51.27 A ATOM 202 O ILE A 30 23.159 11.111 13.473 1.00 50.40A ATOM 203 N TYR A 31 21.878 9.573 14.507 1.00 51.61 A ATOM 204 CA TYR A31 23.018 8.827 15.030 1.00 52.03 A ATOM 205 CB TYR A 31 22.544 7.67315.918 1.00 54.81 A ATOM 206 CG TYR A 31 23.178 7.688 17.296 1.00 58.36A ATOM 207 CD1 TYR A 31 22.785 8.624 18.253 1.00 60.03 A ATOM 208 CE1TYR A 31 23.381 8.666 19.514 1.00 61.40 A ATOM 209 CD2 TYR A 31 24.1896.788 17.632 1.00 59.71 A ATOM 210 CE2 TYR A 31 24.795 6.821 18.891 1.0061.71 A ATOM 211 CZ TYR A 31 24.384 7.762 19.826 1.00 62.61 A ATOM 212OH TYR A 31 24.966 7.797 21.074 1.00 63.90 A ATOM 213 C TYR A 31 23.9958.295 13.989 1.00 50.42 A ATOM 214 O TYR A 31 25.206 8.452 14.141 1.0050.96 A ATOM 215 N PRO A 32 23.490 7.651 12.921 1.00 49.99 A ATOM 216 CDPRO A 32 22.091 7.335 12.584 1.00 49.46 A ATOM 217 CA PRO A 32 24.4017.123 11.898 1.00 49.18 A ATOM 218 CB PRO A 32 23.449 6.566 10.841 1.0048.74 A ATOM 219 CG PRO A 32 22.256 6.163 11.646 1.00 49.30 A ATOM 220 CPRO A 32 25.297 8.222 11.339 1.00 48.53 A ATOM 221 O PRO A 32 26.4587.984 10.993 1.00 48.53 A ATOM 222 N LEU A 33 24.750 9.430 11.267 1.0047.26 A ATOM 223 CA LEU A 33 25.486 10.570 10.745 1.00 47.08 A ATOM 224CB LEU A 33 24.505 11.679 10.356 1.00 46.08 A ATOM 225 CG LEU A 3325.021 12.774 9.422 1.00 45.61 A ATOM 226 CD1 LEU A 33 25.592 12.1528.156 1.00 44.68 A ATOM 227 CD2 LEU A 33 23.875 13.720 9.080 1.00 45.94A ATOM 228 C LEU A 33 26.517 11.095 11.745 1.00 46.98 A ATOM 229 O LEU A33 27.673 11.314 11.392 1.00 46.18 A ATOM 230 N LEU A 34 26.104 11.29012.993 1.00 47.69 A ATOM 231 CA LEU A 34 27.019 11.780 14.021 1.00 49.32A ATOM 232 CB LEU A 34 26.276 11.978 15.348 1.00 49.84 A ATOM 233 CG LEUA 34 25.193 13.061 15.415 1.00 50.05 A ATOM 234 CD1 LEU A 34 24.54513.051 16.786 1.00 49.62 A ATOM 235 CD2 LEU A 34 25.806 14.420 15.1361.00 50.80 A ATOM 236 C LEU A 34 28.202 10.825 14.229 1.00 49.93 A ATOM237 O LEU A 34 29.333 11.262 14.445 1.00 50.30 A ATOM 238 N SER A 3527.936 9.524 14.167 1.00 50.31 A ATOM 239 CA SER A 35 28.984 8.52514.344 1.00 50.96 A ATOM 240 CB SER A 35 28.395 7.114 14.318 1.00 51.84A ATOM 241 OG SER A 35 27.562 6.890 15.442 1.00 54.71 A ATOM 242 C SER A35 30.022 8.652 13.243 1.00 51.29 A ATOM 243 O SER A 35 31.220 8.73813.512 1.00 51.44 A ATOM 244 N THR A 36 29.554 8.669 12.000 1.00 50.28 AATOM 245 CA THR A 36 30.444 8.782 10.855 1.00 49.52 A ATOM 246 CB THR A36 29.645 8.980 9.559 1.00 49.26 A ATOM 247 OG1 THR A 36 28.690 7.9239.425 1.00 49.50 A ATOM 248 CG2 THR A 36 30.568 8.968 8.358 1.00 49.15 AATOM 249 C THR A 36 31.410 9.950 11.025 1.00 49.71 A ATOM 250 O THR A 3632.552 9.893 10.571 1.00 50.28 A ATOM 251 N PHE A 37 30.956 11.00911.689 1.00 49.29 A ATOM 252 CA PHE A 37 31.807 12.174 11.891 1.00 48.84A ATOM 253 CB PHE A 37 31.146 13.416 11.284 1.00 46.61 A ATOM 254 CG PHEA 37 30.934 13.323 9.799 1.00 43.07 A ATOM 255 CD1 PHE A 37 29.72912.873 9.283 1.00 41.56 A ATOM 256 CD2 PHE A 37 31.957 13.656 8.920 1.0039.81 A ATOM 257 CE1 PHE A 37 29.543 12.754 7.908 1.00 42.57 A ATOM 258CE2 PHE A 37 31.785 13.542 7.549 1.00 41.01 A ATOM 259 CZ PHE A 3730.576 13.090 7.039 1.00 40.00 A ATOM 260 C PHE A 37 32.136 12.42313.359 1.00 50.82 A ATOM 261 O PHE A 37 32.304 13.568 13.781 1.00 50.54A ATOM 262 N GLN A 38 32.246 11.343 14.128 1.00 52.93 A ATOM 263 CA GLNA 38 32.543 11.433 15.557 1.00 54.33 A ATOM 264 CB GLN A 38 32.54310.036 16.180 1.00 55.97 A ATOM 265 CG GLN A 38 33.536 9.088 15.535 1.0058.11 A ATOM 266 CD GLN A 38 33.584 7.741 16.221 1.00 59.72 A ATOM 267OE1 GLN A 38 32.549 7.124 16.483 1.00 60.66 A ATOM 268 NE2 GLN A 3834.790 7.271 16.510 1.00 60.33 A ATOM 269 C GLN A 38 33.871 12.11315.870 1.00 53.93 A ATOM 270 O GLN A 38 33.990 12.818 16.868 1.00 53.58A ATOM 271 N ASP A 39 34.868 11.901 15.022 1.00 53.91 A ATOM 272 CA ASPA 39 36.178 12.495 15.252 1.00 54.34 A ATOM 273 CB ASP A 39 37.19911.935 14.259 1.00 55.58 A ATOM 274 CG ASP A 39 37.421 10.443 14.4311.00 57.29 A ATOM 275 OD1 ASP A 39 37.553 9.990 15.587 1.00 57.11 A ATOM276 OD2 ASP A 39 37.475 9.724 13.409 1.00 58.78 A ATOM 277 C ASP A 3936.188 14.019 15.186 1.00 54.75 A ATOM 278 O ASP A 39 37.163 14.65315.594 1.00 53.65 A ATOM 279 N THR A 40 35.104 14.608 14.686 1.00 54.95A ATOM 280 CA THR A 40 35.019 16.061 14.569 1.00 55.03 A ATOM 281 CB THRA 40 34.192 16.487 13.336 1.00 53.61 A ATOM 282 OG1 THR A 40 32.82516.109 13.526 1.00 51.16 A ATOM 283 CG2 THR A 40 34.720 15.833 12.0801.00 52.82 A ATOM 284 C THR A 40 34.366 16.707 15.783 1.00 56.50 A ATOM285 O THR A 40 34.613 17.876 16.082 1.00 57.15 A ATOM 286 N LEU A 4133.536 15.937 16.478 1.00 57.42 A ATOM 287 CA LEU A 41 32.803 16.43217.639 1.00 58.80 A ATOM 288 CB LEU A 41 31.702 15.430 18.004 1.00 57.63A ATOM 289 CG LEU A 41 30.761 15.083 16.844 1.00 56.86 A ATOM 290 CD1LEU A 41 29.770 14.018 17.277 1.00 55.61 A ATOM 291 CD2 LEU A 41 30.03916.341 16.378 1.00 56.19 A ATOM 292 C LEU A 41 33.647 16.751 18.871 1.0060.36 A ATOM 293 O LEU A 41 33.108 17.030 19.942 1.00 60.30 A ATOM 294 NVAL A 42 34.967 16.725 18.719 1.00 62.05 A ATOM 295 CA VAL A 42 35.85917.015 19.834 1.00 63.74 A ATOM 296 CB VAL A 42 37.280 16.494 19.5621.00 64.26 A ATOM 297 CG1 VAL A 42 38.190 16.837 20.733 1.00 65.42 AATOM 298 CG2 VAL A 42 37.243 14.990 19.333 1.00 64.88 A ATOM 299 C VAL A42 35.942 18.510 20.126 1.00 64.12 A ATOM 300 O VAL A 42 36.493 19.27719.332 1.00 64.30 A ATOM 301 N GLU A 43 35.398 18.910 21.273 1.00 64.21A ATOM 302 CA GLU A 43 35.403 20.308 21.694 1.00 64.09 A ATOM 303 CB GLUA 43 35.025 20.410 23.175 1.00 65.49 A ATOM 304 CG GLU A 43 33.62119.924 23.503 1.00 68.00 A ATOM 305 CD GLU A 43 33.338 19.927 24.9971.00 69.13 A ATOM 306 OE1 GLU A 43 34.017 19.180 25.733 1.00 68.77 AATOM 307 OE2 GLU A 43 32.440 20.677 25.436 1.00 70.84 A ATOM 308 C GLU A43 36.770 20.955 21.473 1.00 63.27 A ATOM 309 O GLU A 43 37.795 20.42421.901 1.00 63.43 A ATOM 310 N GLY A 44 36.778 22.102 20.802 1.00 61.60A ATOM 311 CA GLY A 44 38.027 22.792 20.543 1.00 59.61 A ATOM 312 C GLYA 44 38.003 23.516 19.215 1.00 58.39 A ATOM 313 O GLY A 44 37.805 24.73019.161 1.00 59.02 A ATOM 314 N GLY A 45 38.219 22.770 18.138 1.00 56.07A ATOM 315 CA GLY A 45 38.201 23.362 16.816 1.00 54.08 A ATOM 316 C GLYA 45 36.946 22.906 16.101 1.00 52.48 A ATOM 317 O GLY A 45 36.914 22.81714.879 1.00 52.24 A ATOM 318 N SER A 46 35.911 22.613 16.883 1.00 51.11A ATOM 319 CA SER A 46 34.633 22.152 16.358 1.00 48.42 A ATOM 320 CB SERA 46 34.175 20.912 17.122 1.00 48.98 A ATOM 321 OG SER A 46 32.80120.655 16.893 1.00 49.29 A ATOM 322 C SER A 46 33.531 23.203 16.423 1.0047.59 A ATOM 323 O SER A 46 33.421 23.960 17.391 1.00 46.05 A ATOM 324 NVAL A 47 32.711 23.243 15.380 1.00 45.49 A ATOM 325 CA VAL A 47 31.60524.181 15.326 1.00 42.26 A ATOM 326 CB VAL A 47 31.764 25.186 14.1641.00 41.88 A ATOM 327 CG1 VAL A 47 30.602 26.174 14.167 1.00 41.99 AATOM 328 CG2 VAL A 47 33.079 25.937 14.295 1.00 41.85 A ATOM 329 C VAL A47 30.323 23.390 15.129 1.00 41.67 A ATOM 330 O VAL A 47 30.199 22.63114.171 1.00 42.19 A ATOM 331 N VAL A 48 29.388 23.536 16.059 1.00 40.43A ATOM 332 CA VAL A 48 28.111 22.849 15.962 1.00 39.58 A ATOM 333 CB VALA 48 27.974 21.711 16.995 1.00 40.32 A ATOM 334 CG1 VAL A 48 26.52721.222 17.041 1.00 39.82 A ATOM 335 CG2 VAL A 48 28.889 20.553 16.6191.00 40.14 A ATOM 336 C VAL A 48 27.012 23.869 16.197 1.00 39.83 A ATOM337 O VAL A 48 27.061 24.636 17.159 1.00 39.49 A ATOM 338 N VAL A 4926.023 23.874 15.308 1.00 39.22 A ATOM 339 CA VAL A 49 24.914 24.80815.393 1.00 38.77 A ATOM 340 CB VAL A 49 24.974 25.838 14.235 1.00 38.65A ATOM 341 CG1 VAL A 49 24.027 26.994 14.514 1.00 39.77 A ATOM 342 CG2VAL A 49 26.394 26.331 14.047 1.00 37.33 A ATOM 343 C VAL A 49 23.57124.092 15.314 1.00 39.16 A ATOM 344 O VAL A 49 23.427 23.091 14.611 1.0039.92 A ATOM 345 N PHE A 50 22.596 24.601 16.057 1.00 38.97 A ATOM 346CA PHE A 50 21.243 24.061 16.038 1.00 39.87 A ATOM 347 CB PHE A 5020.834 23.575 17.425 1.00 39.82 A ATOM 348 CG PHE A 50 21.720 22.49317.971 1.00 41.79 A ATOM 349 CD1 PHE A 50 22.076 21.400 17.182 1.0042.66 A ATOM 350 CD2 PHE A 50 22.167 22.544 19.288 1.00 41.26 A ATOM 351CE1 PHE A 50 22.864 20.368 17.699 1.00 43.88 A ATOM 352 CE2 PHE A 5022.953 21.520 19.814 1.00 42.58 A ATOM 353 CZ PHE A 50 23.302 20.42919.019 1.00 41.73 A ATOM 354 C PHE A 50 20.380 25.242 15.598 1.00 39.56A ATOM 355 O PHE A 50 20.261 26.227 16.320 1.00 38.63 A ATOM 356 N SER A51 19.772 25.139 14.419 1.00 39.59 A ATOM 357 CA SER A 51 19.002 26.25613.892 1.00 38.98 A ATOM 358 CB SER A 51 19.631 26.692 12.571 1.00 39.41A ATOM 359 OG SER A 51 21.022 26.893 12.739 1.00 40.57 A ATOM 360 C SERA 51 17.500 26.086 13.706 1.00 39.41 A ATOM 361 O SER A 51 17.018 25.03713.283 1.00 38.55 A ATOM 362 N MET A 52 16.778 27.163 14.006 1.00 39.56A ATOM 363 CA MET A 52 15.326 27.203 13.904 1.00 41.09 A ATOM 364 CB META 52 14.721 27.612 15.248 1.00 42.98 A ATOM 365 CG MET A 52 14.95229.080 15.553 1.00 44.79 A ATOM 366 SD MET A 52 14.137 29.694 17.0321.00 53.36 A ATOM 367 CE MET A 52 12.461 29.878 16.420 1.00 50.19 A ATOM368 C MET A 52 14.948 28.254 12.867 1.00 39.86 A ATOM 369 O MET A 5215.736 29.144 12.577 1.00 37.96 A ATOM 370 N ALA A 53 13.735 28.15512.329 1.00 41.96 A ATOM 371 CA ALA A 53 13.240 29.115 11.344 1.00 43.31A ATOM 372 CB ALA A 53 13.443 28.583 9.930 1.00 43.60 A ATOM 373 C ALA A53 11.757 29.373 11.605 1.00 44.82 A ATOM 374 O ALA A 53 11.035 28.48612.058 1.00 45.88 A ATOM 375 N SER A 54 11.309 30.588 11.314 1.00 45.31A ATOM 376 CA SER A 54 9.920 30.975 11.536 1.00 46.31 A ATOM 377 CB SERA 54 9.824 32.490 11.704 1.00 44.93 A ATOM 378 OG SER A 54 9.947 33.12610.439 1.00 42.54 A ATOM 379 C SER A 54 8.992 30.569 10.402 1.00 47.54 AATOM 380 O SER A 54 9.417 30.002 9.398 1.00 47.54 A ATOM 381 N GLY A 557.713 30.889 10.583 1.00 50.46 A ATOM 382 CA GLY A 55 6.701 30.596 9.5851.00 52.05 A ATOM 383 C GLY A 55 6.526 29.136 9.219 1.00 52.73 A ATOM384 O GLY A 55 6.502 28.256 10.081 1.00 52.54 A ATOM 385 N ARG A 566.399 28.887 7.922 1.00 53.50 A ATOM 386 CA ARG A 56 6.206 27.539 7.4081.00 54.78 A ATOM 387 CB ARG A 56 5.885 27.590 5.911 1.00 55.54 A ATOM388 CG ARG A 56 7.112 27.702 5.021 1.00 55.10 A ATOM 389 CD ARG A 566.731 27.742 3.551 1.00 56.42 A ATOM 390 NE ARG A 56 7.892 27.536 2.6901.00 58.46 A ATOM 391 CZ ARG A 56 7.858 27.590 1.363 1.00 59.31 A ATOM392 NH1 ARG A 56 6.718 27.849 0.737 1.00 59.88 A ATOM 393 NH2 ARG A 568.961 27.375 0.658 1.00 59.39 A ATOM 394 C ARG A 56 7.438 26.666 7.6261.00 54.74 A ATOM 395 O ARG A 56 7.404 25.464 7.359 1.00 54.18 A ATOM396 N HIS A 57 8.522 27.270 8.105 1.00 54.51 A ATOM 397 CA HIS A 579.759 26.525 8.335 1.00 54.29 A ATOM 398 CB HIS A 57 10.964 27.302 7.7791.00 55.16 A ATOM 399 CG HIS A 57 10.719 27.936 6.444 1.00 56.03 A ATOM400 CD2 HIS A 57 11.050 27.537 5.193 1.00 56.33 A ATOM 401 ND1 HIS A 5710.032 29.122 6.298 1.00 56.66 A ATOM 402 CE1 HIS A 57 9.948 29.4265.014 1.00 56.21 A ATOM 403 NE2 HIS A 57 10.557 28.480 4.322 1.00 56.40A ATOM 404 C HIS A 57 9.986 26.266 9.822 1.00 53.88 A ATOM 405 O HIS A57 11.051 25.791 10.220 1.00 53.33 A ATOM 406 N SER A 58 8.980 26.55910.639 1.00 53.09 A ATOM 407 CA SER A 58 9.108 26.402 12.085 1.00 52.87A ATOM 408 CB SER A 58 8.133 27.346 12.794 1.00 53.36 A ATOM 409 OG SERA 58 6.792 27.058 12.437 1.00 54.09 A ATOM 410 C SER A 58 8.967 25.00912.689 1.00 51.52 A ATOM 411 O SER A 58 9.114 24.859 13.901 1.00 52.42 AATOM 412 N THR A 59 8.696 23.995 11.874 1.00 49.99 A ATOM 413 CA THR A59 8.540 22.644 12.408 1.00 49.61 A ATOM 414 CB THR A 59 7.321 21.92611.805 1.00 50.26 A ATOM 415 OG1 THR A 59 7.565 21.660 10.418 1.00 52.05A ATOM 416 CG2 THR A 59 6.070 22.778 11.955 1.00 49.84 A ATOM 417 C THRA 59 9.750 21.755 12.168 1.00 48.80 A ATOM 418 O THR A 59 9.627 20.53412.141 1.00 50.12 A ATOM 419 N GLU A 60 10.917 22.359 11.996 1.00 47.87A ATOM 420 CA GLU A 60 12.127 21.583 11.760 1.00 48.24 A ATOM 421 CB GLUA 60 12.428 21.520 10.260 1.00 49.44 A ATOM 422 CG GLU A 60 11.63820.460 9.520 1.00 53.29 A ATOM 423 CD GLU A 60 11.520 20.742 8.030 1.0056.50 A ATOM 424 OE1 GLU A 60 12.551 21.045 7.391 1.00 55.69 A ATOM 425OE2 GLU A 60 10.389 20.657 7.497 1.00 59.38 A ATOM 426 C GLU A 60 13.32822.148 12.498 1.00 46.12 A ATOM 427 O GLU A 60 13.344 23.319 12.886 1.0046.85 A ATOM 428 N LEU A 61 14.328 21.301 12.708 1.00 43.44 A ATOM 429CA LEU A 61 15.544 21.737 13.374 1.00 41.98 A ATOM 430 CB LEU A 6115.603 21.245 14.822 1.00 41.96 A ATOM 431 CG LEU A 61 16.830 21.73615.603 1.00 40.28 A ATOM 432 CD1 LEU A 61 16.748 23.234 15.798 1.0041.49 A ATOM 433 CD2 LEU A 61 16.899 21.043 16.957 1.00 43.82 A ATOM 434C LEU A 61 16.748 21.216 12.613 1.00 40.23 A ATOM 435 O LEU A 61 16.95120.005 12.487 1.00 38.93 A ATOM 436 N ASP A 62 17.535 22.153 12.100 1.0039.30 A ATOM 437 CA ASP A 62 18.736 21.840 11.348 1.00 38.26 A ATOM 438CB ASP A 62 18.972 22.893 10.263 1.00 37.02 A ATOM 439 CG ASP A 6218.188 22.617 8.993 1.00 38.78 A ATOM 440 OD1 ASP A 62 17.534 21.5578.915 1.00 38.35 A ATOM 441 OD2 ASP A 62 18.234 23.459 8.069 1.00 35.53A ATOM 442 C ASP A 62 19.948 21.817 12.261 1.00 37.32 A ATOM 443 O ASP A62 20.004 22.555 13.251 1.00 38.00 A ATOM 444 N PHE A 63 20.905 20.95611.938 1.00 36.34 A ATOM 445 CA PHE A 63 22.142 20.894 12.692 1.00 36.73A ATOM 446 CB PHE A 63 22.235 19.624 13.552 1.00 38.45 A ATOM 447 CG PHEA 63 22.102 18.336 12.790 1.00 40.09 A ATOM 448 CD1 PHE A 63 20.84917.814 12.491 1.00 41.21 A ATOM 449 CD2 PHE A 63 23.230 17.620 12.4131.00 38.85 A ATOM 450 CE1 PHE A 63 20.722 16.596 11.830 1.00 41.80 AATOM 451 CE2 PHE A 63 23.117 16.405 11.753 1.00 40.48 A ATOM 452 CZ PHEA 63 21.857 15.889 11.460 1.00 41.84 A ATOM 453 C PHE A 63 23.297 20.96111.709 1.00 36.67 A ATOM 454 O PHE A 63 23.337 20.215 10.724 1.00 37.63A ATOM 455 N SER A 64 24.217 21.884 11.966 1.00 37.08 A ATOM 456 CA SERA 64 25.389 22.078 11.123 1.00 37.11 A ATOM 457 CB SER A 64 25.53323.550 10.742 1.00 37.02 A ATOM 458 OG SER A 64 24.347 24.054 10.1561.00 37.59 A ATOM 459 C SER A 64 26.617 21.641 11.908 1.00 37.78 A ATOM460 O SER A 64 26.776 22.004 13.070 1.00 38.14 A ATOM 461 N ILE A 6527.486 20.869 11.269 1.00 37.84 A ATOM 462 CA ILE A 65 28.687 20.37511.931 1.00 38.20 A ATOM 463 CB ILE A 65 28.517 18.881 12.298 1.00 38.18A ATOM 464 CG2 ILE A 65 29.813 18.319 12.870 1.00 39.41 A ATOM 465 CG1ILE A 65 27.372 18.732 13.306 1.00 37.70 A ATOM 466 CD1 ILE A 65 26.94717.304 13.552 1.00 38.47 A ATOM 467 C ILE A 65 29.918 20.549 11.053 1.0038.20 A ATOM 468 O ILE A 65 29.936 20.120 9.903 1.00 38.89 A ATOM 469 NSER A 66 30.947 21.184 11.598 1.00 38.03 A ATOM 470 CA SER A 66 32.17821.396 10.848 1.00 38.52 A ATOM 471 CB SER A 66 33.091 22.367 11.5991.00 37.05 A ATOM 472 OG SER A 66 33.302 21.907 12.920 1.00 39.76 A ATOM473 C SER A 66 32.897 20.067 10.652 1.00 38.47 A ATOM 474 O SER A 6632.773 19.162 11.474 1.00 39.22 A ATOM 475 N VAL A 67 33.635 19.9549.553 1.00 37.19 A ATOM 476 CA VAL A 67 34.393 18.745 9.242 1.00 36.16 AATOM 477 CB VAL A 67 33.639 17.821 8.251 1.00 36.78 A ATOM 478 CG1 VAL A67 34.466 16.561 7.977 1.00 34.33 A ATOM 479 CG2 VAL A 67 32.277 17.4398.816 1.00 33.97 A ATOM 480 C VAL A 67 35.715 19.174 8.604 1.00 36.50 AATOM 481 O VAL A 67 35.739 19.707 7.496 1.00 34.41 A ATOM 482 N PRO A 6836.834 18.970 9.315 1.00 38.13 A ATOM 483 CD PRO A 68 36.945 18.35610.650 1.00 38.47 A ATOM 484 CA PRO A 68 38.156 19.346 8.795 1.00 38.92A ATOM 485 CB PRO A 68 39.104 18.943 9.927 1.00 39.16 A ATOM 486 CG PROA 68 38.355 17.833 10.630 1.00 38.66 A ATOM 487 C PRO A 68 38.469 18.6387.482 1.00 38.57 A ATOM 488 O PRO A 68 38.045 17.506 7.274 1.00 39.03 AATOM 489 N THR A 69 39.200 19.309 6.598 1.00 40.67 A ATOM 490 CA THR A69 39.542 18.736 5.297 1.00 41.89 A ATOM 491 CB THR A 69 40.363 19.7194.449 1.00 41.81 A ATOM 492 OG1 THR A 69 41.533 20.109 5.174 1.00 45.53A ATOM 493 CG2 THR A 69 39.548 20.949 4.115 1.00 41.08 A ATOM 494 C THRA 69 40.335 17.443 5.420 1.00 42.77 A ATOM 495 O THR A 69 40.345 16.6194.499 1.00 40.61 A ATOM 496 N SER A 70 40.992 17.270 6.563 1.00 43.04 AATOM 497 CA SER A 70 41.802 16.083 6.810 1.00 44.22 A ATOM 498 CB SER A70 42.565 16.231 8.133 1.00 44.13 A ATOM 499 OG SER A 70 41.679 16.2549.239 1.00 42.16 A ATOM 500 C SER A 70 40.934 14.833 6.854 1.00 44.81 AATOM 501 O SER A 70 41.387 13.734 6.531 1.00 45.00 A ATOM 502 N HIS A 7139.679 15.000 7.250 1.00 44.99 A ATOM 503 CA HIS A 71 38.779 13.8667.328 1.00 45.50 A ATOM 504 CB HIS A 71 37.595 14.215 8.230 1.00 50.25 AATOM 505 CG HIS A 71 37.970 14.374 9.671 1.00 54.93 A ATOM 506 CD2 HIS A71 39.181 14.404 10.279 1.00 56.60 A ATOM 507 ND1 HIS A 71 37.036 14.52510.674 1.00 56.61 A ATOM 508 CE1 HIS A 71 37.656 14.640 11.837 1.0057.31 A ATOM 509 NE2 HIS A 71 38.957 14.570 11.625 1.00 57.50 A ATOM 510C HIS A 71 38.313 13.397 5.948 1.00 44.27 A ATOM 511 O HIS A 71 37.77812.299 5.803 1.00 42.77 A ATOM 512 N GLY A 72 38.539 14.226 4.935 1.0042.93 A ATOM 513 CA GLY A 72 38.151 13.868 3.582 1.00 42.01 A ATOM 514 CGLY A 72 36.825 14.454 3.136 1.00 40.53 A ATOM 515 O GLY A 72 36.04614.934 3.954 1.00 40.70 A ATOM 516 N ASP A 73 36.581 14.423 1.828 1.0039.45 A ATOM 517 CA ASP A 73 35.345 14.931 1.238 1.00 39.65 A ATOM 518CB ASP A 73 35.309 14.568 −0.251 1.00 39.15 A ATOM 519 CG ASP A 7334.036 15.025 −0.942 1.00 37.87 A ATOM 520 OD1 ASP A 73 32.947 14.913−0.343 1.00 39.76 A ATOM 521 OD2 ASP A 73 34.122 15.485 −2.097 1.0040.00 A ATOM 522 C ASP A 73 34.162 14.280 1.957 1.00 37.48 A ATOM 523 OASP A 73 33.993 13.066 1.895 1.00 38.41 A ATOM 524 N PRO A 74 33.33515.079 2.652 1.00 36.55 A ATOM 525 CD PRO A 74 33.529 16.507 2.958 1.0037.15 A ATOM 526 CA PRO A 74 32.175 14.544 3.378 1.00 34.86 A ATOM 527CB PRO A 74 31.752 15.714 4.268 1.00 34.11 A ATOM 528 CG PRO A 74 32.17016.906 3.466 1.00 36.59 A ATOM 529 C PRO A 74 31.027 14.004 2.518 1.0033.80 A ATOM 530 O PRO A 74 30.274 13.156 2.982 1.00 33.04 A ATOM 531 NTYR A 75 30.867 14.494 1.289 1.00 32.38 A ATOM 532 CA TYR A 75 29.80113.957 0.447 1.00 32.91 A ATOM 533 CB TYR A 75 29.530 14.819 −0.789 1.0032.39 A ATOM 534 CG TYR A 75 28.334 14.320 −1.600 1.00 32.65 A ATOM 535CD1 TYR A 75 27.086 14.159 −1.002 1.00 32.34 A ATOM 536 CE1 TYR A 7525.978 13.752 −1.731 1.00 32.78 A ATOM 537 CD2 TYR A 75 28.442 14.047−2.966 1.00 35.34 A ATOM 538 CE2 TYR A 75 27.321 13.632 −3.714 1.0033.58 A ATOM 539 CZ TYR A 75 26.098 13.494 −3.084 1.00 33.86 A ATOM 540OH TYR A 75 24.972 13.132 −3.797 1.00 33.63 A ATOM 541 C TYR A 75 30.25612.580 −0.007 1.00 33.77 A ATOM 542 O TYR A 75 29.464 11.641 −0.055 1.0033.18 A ATOM 543 N ALA A 76 31.540 12.470 −0.346 1.00 34.39 A ATOM 544CA ALA A 76 32.098 11.187 −0.772 1.00 35.15 A ATOM 545 CB ALA A 7633.595 11.319 −1.071 1.00 34.26 A ATOM 546 C ALA A 76 31.885 10.1990.362 1.00 34.76 A ATOM 547 O ALA A 76 31.608 9.023 0.136 1.00 33.20 AATOM 548 N THR A 77 32.005 10.695 1.588 1.00 34.77 A ATOM 549 CA THR A77 31.824 9.858 2.767 1.00 36.05 A ATOM 550 CB THR A 77 32.317 10.5844.034 1.00 36.33 A ATOM 551 OG1 THR A 77 33.740 10.719 3.968 1.00 37.57A ATOM 552 CG2 THR A 77 31.941 9.798 5.289 1.00 36.53 A ATOM 553 C THR A77 30.385 9.390 2.999 1.00 37.01 A ATOM 554 O THR A 77 30.152 8.2033.263 1.00 38.11 A ATOM 555 N VAL A 78 29.420 10.303 2.911 1.00 36.63 AATOM 556 CA VAL A 78 28.027 9.923 3.138 1.00 36.66 A ATOM 557 CB VAL A78 27.087 11.164 3.268 1.00 36.95 A ATOM 558 CG1 VAL A 78 27.499 11.9984.468 1.00 36.05 A ATOM 559 CG2 VAL A 78 27.124 12.002 1.999 1.00 36.59A ATOM 560 C VAL A 78 27.480 9.003 2.053 1.00 35.80 A ATOM 561 O VAL A78 26.614 8.176 2.325 1.00 35.84 A ATOM 562 N VAL A 79 27.972 9.1350.826 1.00 35.93 A ATOM 563 CA VAL A 79 27.485 8.269 −0.241 1.00 36.96 AATOM 564 CB VAL A 79 27.857 8.808 −1.641 1.00 36.08 A ATOM 565 CG1 VAL A79 27.323 7.859 −2.712 1.00 34.68 A ATOM 566 CG2 VAL A 79 27.278 10.197−1.844 1.00 36.45 A ATOM 567 C VAL A 79 28.091 6.872 −0.064 1.00 37.92 AATOM 568 O VAL A 79 27.384 5.867 −0.111 1.00 38.87 A ATOM 569 N GLU A 8029.402 6.823 0.155 1.00 38.87 A ATOM 570 CA GLU A 80 30.106 5.562 0.3451.00 40.08 A ATOM 571 CB GLU A 80 31.589 5.809 0.592 1.00 42.04 A ATOM572 CG GLU A 80 32.435 5.911 −0.653 1.00 47.22 A ATOM 573 CD GLU A 8033.913 5.949 −0.316 1.00 50.49 A ATOM 574 OE1 GLU A 80 34.379 5.0270.396 1.00 51.53 A ATOM 575 OE2 GLU A 80 34.603 6.895 −0.760 1.00 52.54A ATOM 576 C GLU A 80 29.579 4.732 1.502 1.00 40.32 A ATOM 577 O GLU A80 29.569 3.504 1.436 1.00 39.33 A ATOM 578 N LYS A 81 29.158 5.4012.567 1.00 40.75 A ATOM 579 CA LYS A 81 28.658 4.701 3.743 1.00 42.50 AATOM 580 CB LYS A 81 28.979 5.515 4.999 1.00 43.97 A ATOM 581 CG LYS A81 28.806 4.473 6.294 1.00 47.62 A ATOM 582 CD LYS A 81 29.508 5.4377.448 1.00 48.42 A ATOM 583 CE LYS A 81 29.424 4.611 8.722 1.00 49.73 AATOM 584 NZ LYS A 81 30.230 5.215 9.816 1.00 50.46 A ATOM 585 C LYS A 8127.160 4.393 3.659 1.00 42.43 A ATOM 586 O LYS A 81 26.583 3.794 4.5691.00 43.82 A ATOM 587 N GLY A 82 26.534 4.800 2.561 1.00 41.47 A ATOM588 CA GLY A 82 25.120 4.530 2.380 1.00 40.57 A ATOM 589 C GLY A 8224.167 5.412 3.165 1.00 39.41 A ATOM 590 O GLY A 82 23.027 5.017 3.4101.00 39.15 A ATOM 591 N LEU A 83 24.617 6.606 3.551 1.00 38.67 A ATOM592 CA LEU A 83 23.776 7.530 4.310 1.00 36.40 A ATOM 593 CB LEU A 8324.625 8.331 5.296 1.00 37.66 A ATOM 594 CG LEU A 83 25.301 7.524 6.4091.00 39.03 A ATOM 595 CD1 LEU A 83 26.040 8.469 7.328 1.00 39.46 A ATOM596 CD2 LEU A 83 24.260 6.734 7.203 1.00 38.72 A ATOM 597 C LEU A 8322.999 8.481 3.407 1.00 36.46 A ATOM 598 O LEU A 83 22.058 9.144 3.8451.00 38.34 A ATOM 599 N PHE A 84 23.397 8.569 2.145 1.00 33.71 A ATOM600 CA PHE A 84 22.695 9.427 1.206 1.00 34.20 A ATOM 601 CB PHE A 8423.172 10.885 1.305 1.00 35.21 A ATOM 602 CG PHE A 84 23.307 11.8400.541 1.00 33.04 A ATOM 603 CD1 PHE A 84 21.109 12.290 1.080 1.00 33.86A ATOM 604 CD2 PHE A 84 22.636 12.211 −0.758 1.00 32.80 A ATOM 605 CE1PHE A 84 20.247 13.088 0.335 1.00 33.51 A ATOM 606 CE2 PHE A 84 21.78013.010 −1.513 1.00 32.34 A ATOM 607 CZ PHE A 84 20.583 13.449 −0.9701.00 33.64 A ATOM 608 C PHE A 84 22.952 8.909 −0.199 1.00 34.14 A ATOM609 O PHE A 84 24.055 8.456 −0.502 1.00 32.87 A ATOM 610 N PRO A 8521.936 8.965 −1.075 1.00 35.40 A ATOM 611 CD PRO A 85 20.507 9.187−0.785 1.00 35.87 A ATOM 612 CA PRO A 85 22.115 8.481 −2.446 1.00 36.99A ATOM 613 CB PRO A 85 20.743 7.912 −2.782 1.00 36.41 A ATOM 614 CG PROA 85 19.836 8.885 −2.131 1.00 36.37 A ATOM 615 C PRO A 85 22.542 9.560−3.432 1.00 38.58 A ATOM 616 O PRO A 85 22.088 10.697 −3.343 1.00 37.08A ATOM 617 N ALA A 86 23.423 9.204 −4.365 1.00 41.13 A ATOM 618 CA ALA A86 23.854 10.154 −5.378 1.00 43.66 A ATOM 619 CB ALA A 86 24.889 9.528−6.295 1.00 43.21 A ATOM 620 C ALA A 86 22.605 10.524 −6.171 1.00 45.42A ATOM 621 O ALA A 86 21.847 9.649 −6.600 1.00 45.70 A ATOM 622 N THR A87 22.389 11.821 −6.355 1.00 46.96 A ATOM 623 CA THR A 87 21.222 12.308−7.076 1.00 47.91 A ATOM 624 CB THR A 87 21.011 13.810 −6.820 1.00 49.25A ATOM 625 OG1 THR A 87 22.142 14.537 −7.318 1.00 50.27 A ATOM 626 CG2THR A 87 20.850 14.084 −5.323 1.00 48.15 A ATOM 627 C THR A 87 21.30812.094 −8.581 1.00 48.08 A ATOM 628 O THR A 87 20.289 12.120 −9.266 1.0048.96 A ATOM 629 N GLY A 88 22.515 11.868 −9.088 1.00 47.81 A ATOM 630CA GLY A 88 22.701 11.692 −10.520 1.00 47.20 A ATOM 631 C GLY A 8822.574 13.042 −11.212 1.00 47.00 A ATOM 632 O GLY A 88 22.365 13.131−12.424 1.00 47.22 A ATOM 633 N HIS A 89 22.720 14.099 −10.419 1.0045.03 A ATOM 634 CA HIS A 89 22.601 15.482 −10.877 1.00 44.02 A ATOM 635CB HIS A 89 21.499 16.149 −10.033 1.00 45.14 A ATOM 636 CG HIS A 8921.222 17.577 −10.380 1.00 47.25 A ATOM 637 CD2 HIS A 89 20.207 18.143−11.075 1.00 47.80 A ATOM 638 ND1 HIS A 89 22.043 18.612 −9.987 1.0046.81 A ATOM 639 CE1 HIS A 89 21.545 19.755 −10.426 1.00 46.54 A ATOM640 NE2 HIS A 89 20.432 19.499 −11.089 1.00 46.38 A ATOM 641 C HIS A 8923.962 16.183 −10.701 1.00 42.11 A ATOM 642 O HIS A 89 24.302 15.715−9.932 1.00 41.05 A ATOM 643 N PRO A 90 24.208 17.292 −11.425 1.00 40.53A ATOM 644 CD PRO A 90 23.425 17.878 −12.529 1.00 41.84 A ATOM 645 CAPRO A 90 25.492 17.990 −11.285 1.00 39.95 A ATOM 646 CB PRO A 90 25.28219.254 −12.103 1.00 39.62 A ATOM 647 CG PRO A 90 24.447 18.759 −13.2241.00 42.14 A ATOM 648 C PRO A 90 25.879 18.295 −9.836 1.00 38.75 A ATOM649 O PRO A 90 27.062 18.400 −9.508 1.00 36.43 A ATOM 650 N VAL A 9124.884 18.432 −8.967 1.00 37.40 A ATOM 651 CA VAL A 91 25.174 18.729−7.573 1.00 35.42 A ATOM 652 CB VAL A 91 23.863 18.890 −6.751 1.00 32.89A ATOM 653 CG1 VAL A 91 23.089 17.585 −6.708 1.00 29.58 A ATOM 654 CG2VAL A 91 24.188 19.390 −5.348 1.00 33.88 A ATOM 655 C VAL A 91 26.07017.647 −6.973 1.00 36.43 A ATOM 656 O VAL A 91 26.809 17.897 −6.014 1.0035.56 A ATOM 657 N ASP A 92 26.041 16.453 −7.562 1.00 36.81 A ATOM 658CA ASP A 92 26.862 15.352 −7.064 1.00 37.47 A ATOM 659 CB ASP A 9226.513 14.033 −7.764 1.00 39.17 A ATOM 660 CG ASP A 92 25.111 13.547−7.441 1.00 42.34 A ATOM 661 OD1 ASP A 92 24.703 13.605 −6.257 1.0042.11 A ATOM 662 OD2 ASP A 92 24.426 13.094 −8.379 1.00 41.11 A ATOM 663C ASP A 92 28.358 15.600 −7.241 1.00 37.40 A ATOM 664 O ASP A 92 29.17615.030 −6.522 1.00 36.00 A ATOM 665 N ASP A 93 28.714 16.443 −8.200 1.0037.21 A ATOM 666 CA ASP A 93 30.119 16.714 −8.465 1.00 38.02 A ATOM 667CB ASP A 93 30.378 16.662 −9.974 1.00 39.00 A ATOM 668 CG ASP A 9329.861 15.385 −10.511 1.00 42.85 A ATOM 669 OD1 ASP A 93 30.238 14.293−10.139 1.00 43.31 A ATOM 670 OD2 ASP A 93 29.075 15.476 −11.579 1.0045.48 A ATOM 671 C ASP A 93 30.625 18.044 −7.932 1.00 37.00 A ATOM 672 OASP A 93 31.831 18.245 −7.835 1.00 36.56 A ATOM 673 N LEU A 94 29.71018.938 −7.568 1.00 35.71 A ATOM 674 CA LEU A 94 30.104 20.268 −7.1201.00 34.14 A ATOM 675 CB LEU A 94 28.867 21.131 −6.853 1.00 34.32 A ATOM676 CG LEU A 94 29.151 22.629 −6.678 1.00 33.22 A ATOM 677 CD1 LEU A 9430.004 23.160 −7.823 1.00 33.67 A ATOM 678 CD2 LEU A 94 27.839 23.368−6.627 1.00 34.83 A ATOM 679 C LEU A 94 31.054 20.357 −5.931 1.00 33.54A ATOM 680 O LEU A 94 32.010 21.132 −5.971 1.00 32.88 A ATOM 681 N LEU A95 30.814 19.592 −4.874 1.00 33.38 A ATOM 682 CA LEU A 95 31.729 19.687−3.749 1.00 34.21 A ATOM 683 CB LEU A 95 31.309 18.780 −2.589 1.00 31.65A ATOM 684 CG LEU A 95 32.185 18.988 −1.337 1.00 31.56 A ATOM 685 CD1LEU A 95 32.251 20.480 −0.967 1.00 32.11 A ATOM 686 CD2 LEU A 95 31.62018.187 −0.173 1.00 32.42 A ATOM 687 C LEU A 95 33.111 19.300 −4.249 1.0034.32 A ATOM 688 O LEU A 95 34.074 20.036 −4.047 1.00 33.01 A ATOM 689 NALA A 96 33.197 18.158 −4.933 1.00 34.56 A ATOM 690 CA ALA A 96 34.47317.675 −5.457 1.00 35.45 A ATOM 691 CB ALA A 96 34.284 16.303 −6.1041.00 37.29 A ATOM 692 C ALA A 96 35.132 18.641 −6.447 1.00 34.95 A ATOM693 O ALA A 96 36.340 18.899 −6.353 1.00 35.01 A ATOM 694 N ASP A 9734.358 19.175 −7.392 1.00 34.87 A ATOM 695 CA ASP A 97 34.915 20.110−8.377 1.00 34.02 A ATOM 696 CB ASP A 97 33.874 20.486 −9.439 1.00 35.17A ATOM 697 CG ASP A 97 33.636 19.385 −10.457 1.00 36.22 A ATOM 698 OD1ASP A 97 34.319 18.338 −10.405 1.00 37.31 A ATOM 699 OD2 ASP A 97 32.75219.580 −11.318 1.00 35.57 A ATOM 700 C ASP A 97 35.412 21.393 −7.7251.00 34.14 A ATOM 701 O ASP A 97 36.325 22.052 −8.232 1.00 33.33 A ATOM702 N THR A 98 34.791 21.757 −6.609 1.00 34.23 A ATOM 703 CA THR A 9835.155 22.966 −5.885 1.00 34.54 A ATOM 704 CB THR A 98 34.095 23.286−4.795 1.00 33.81 A ATOM 705 OG1 THR A 98 32.868 23.653 −5.433 1.0032.38 A ATOM 706 CG2 THR A 98 34.551 24.430 −3.889 1.00 32.67 A ATOM 707C THR A 98 36.528 22.809 −5.242 1.00 35.23 A ATOM 708 O THR A 98 37.35623.722 −5.282 1.00 36.85 A ATOM 709 N GLN A 99 36.764 21.645 −4.652 1.0036.14 A ATOM 710 CA GLN A 99 38.041 21.361 −4.005 1.00 38.58 A ATOM 711CB GLN A 99 37.919 20.077 −3.174 1.00 40.12 A ATOM 712 CG GLN A 9939.212 19.594 −2.530 1.00 44.11 A ATOM 713 CD GLN A 99 38.962 18.628−1.378 1.00 45.38 A ATOM 714 OE1 GLN A 99 38.046 17.800 −1.428 1.0044.91 A ATOM 715 NE2 GLN A 99 39.784 18.723 −0.336 1.00 45.37 A ATOM 716C GLN A 99 39.121 21.224 −5.079 1.00 39.44 A ATOM 717 O GLN A 99 40.30921.422 −4.819 1.00 39.54 A ATOM 718 N LYS A 100 38.693 20.899 −6.2941.00 39.83 A ATOM 719 CA LYS A 100 39.603 20.754 −7.423 1.00 41.72 AATOM 720 CB LYS A 100 38.913 19.961 −8.541 1.00 43.31 A ATOM 721 CG LYSA 100 39.711 19.870 −9.841 1.00 47.29 A ATOM 722 CD LYS A 100 38.87019.299 −10.986 1.00 49.04 A ATOM 723 CE LYS A 100 39.606 19.396 −12.3271.00 50.86 A ATOM 724 NZ LYS A 100 38.770 18.967 −13.495 1.00 52.06 AATOM 725 C LYS A 100 40.085 22.113 −7.970 1.00 41.64 A ATOM 726 O LYS A100 41.277 22.294 −8.241 1.00 42.73 A ATOM 727 N HIS A 101 39.163 23.062−8.126 1.00 40.66 A ATOM 728 CA HIS A 101 39.494 24.378 −8.667 1.0040.67 A ATOM 729 CB HIS A 101 38.349 24.901 −9.546 1.00 41.87 A ATOM 730CG HIS A 101 38.153 24.138 −10.817 1.00 41.74 A ATOM 731 CD2 HIS A 10138.455 24.453 −12.100 1.00 42.24 A ATOM 732 ND1 HIS A 101 37.587 22.882−10.854 1.00 43.06 A ATOM 733 CE1 HIS A 101 37.547 22.456 −12.104 1.0042.69 A ATOM 734 NE2 HIS A 101 38.069 23.390 −12.880 1.00 42.38 A ATOM735 C HIS A 101 39.835 25.475 −7.664 1.00 40.29 A ATOM 736 O HIS A 10140.307 26.537 −8.065 1.00 41.90 A ATOM 737 N LEU A 102 39.598 25.243−6.376 1.00 39.72 A ATOM 738 CA LEU A 102 39.869 26.279 −5.378 1.0038.61 A ATOM 739 CB LEU A 102 38.589 27.073 −5.068 1.00 37.62 A ATOM 740CG LEU A 102 37.875 27.877 −6.156 1.00 37.33 A ATOM 741 CD1 LEU A 10236.523 28.356 −5.621 1.00 37.68 A ATOM 742 CD2 LEU A 102 38.736 29.062−6.586 1.00 38.82 A ATOM 743 C LEU A 102 40.412 25.735 −4.069 1.00 37.71A ATOM 744 O LEU A 102 40.259 24.558 −3.756 1.00 37.42 A ATOM 745 N PROA 103 41.055 26.603 −3.279 1.00 38.28 A ATOM 746 CD PRO A 103 41.51927.947 −3.660 1.00 38.45 A ATOM 747 CA PRO A 103 41.620 26.209 −1.9881.00 38.75 A ATOM 748 CB PRO A 103 42.559 27.367 −1.647 1.00 39.03 AATOM 749 CG PRO A 103 42.849 28.006 −2.977 1.00 39.90 A ATOM 750 C PRO A103 40.501 26.084 −0.962 1.00 38.05 A ATOM 751 O PRO A 103 39.871 27.079−0.608 1.00 39.88 A ATOM 752 N VAL A 104 40.232 24.867 −0.507 1.00 38.36A ATOM 753 CA VAL A 104 39.196 24.658 0.492 1.00 38.23 A ATOM 754 CB VALA 104 38.300 23.457 0.126 1.00 37.32 A ATOM 755 CG1 VAL A 104 37.28823.195 1.226 1.00 35.34 A ATOM 756 CG2 VAL A 104 37.582 23.748 −1.1901.00 37.51 A ATOM 757 C VAL A 104 39.902 24.426 1.819 1.00 39.33 A ATOM758 O VAL A 104 40.699 23.503 1.963 1.00 39.67 A ATOM 759 N SER A 10539.604 25.284 2.783 1.00 39.48 A ATOM 760 CA SER A 105 40.228 25.2304.098 1.00 38.55 A ATOM 761 CB SER A 105 40.410 26.658 4.613 1.00 37.61A ATOM 762 OG SER A 105 39.157 27.324 4.641 1.00 38.92 A ATOM 763 C SERA 105 39.452 24.421 5.127 1.00 37.52 A ATOM 764 O SER A 105 39.99124.060 6.176 1.00 36.61 A ATOM 765 N MET A 106 38.188 24.132 4.836 1.0036.90 A ATOM 766 CA MET A 106 37.368 23.387 5.773 1.00 35.42 A ATOM 767CB MET A 106 37.114 24.245 7.013 1.00 39.45 A ATOM 768 CG MET A 10636.289 23.574 8.088 1.00 44.17 A ATOM 769 SD MET A 106 36.685 24.1999.727 1.00 52.21 A ATOM 770 CE MET A 106 35.296 25.277 10.026 1.00 49.49A ATOM 771 C MET A 106 36.036 23.000 5.147 1.00 34.14 A ATOM 772 O MET A106 35.596 23.624 4.187 1.00 31.92 A ATOM 773 N PHE A 107 35.413 21.9695.706 1.00 31.68 A ATOM 774 CA PHE A 107 34.116 21.479 5.251 1.00 32.11A ATOM 775 CB PHE A 107 34.186 19.993 4.869 1.00 32.56 A ATOM 776 CG PHEA 107 35.046 19.685 3.674 1.00 32.69 A ATOM 777 CD1 PHE A 107 36.10718.789 3.787 1.00 33.01 A ATOM 778 CD2 PHE A 107 34.756 20.223 2.4251.00 34.08 A ATOM 779 CE1 PHE A 107 36.865 18.429 2.675 1.00 33.52 AATOM 780 CE2 PHE A 107 35.514 19.867 1.296 1.00 35.10 A ATOM 781 CZ PHEA 107 36.565 18.969 1.426 1.00 34.67 A ATOM 782 C PHE A 107 33.10721.581 6.395 1.00 30.99 A ATOM 783 O PHE A 107 33.445 21.943 7.525 1.0030.91 A ATOM 784 N ALA A 108 31.861 21.248 6.082 1.00 31.37 A ATOM 785CA ALA A 108 30.798 21.204 7.071 1.00 29.87 A ATOM 786 CB ALA A 10830.384 22.603 7.494 1.00 29.38 A ATOM 787 C ALA A 108 29.629 20.4576.441 1.00 30.97 A ATOM 788 O ALA A 108 29.518 20.383 5.211 1.00 29.63 AATOM 789 N ILE A 109 28.790 19.855 7.274 1.00 31.61 A ATOM 790 CA ILE A109 27.624 19.156 6.673 1.00 33.76 A ATOM 791 CB ILE A 109 27.718 17.6276.925 1.00 35.00 A ATOM 792 CG2 ILE A 109 28.841 17.086 6.047 1.00 34.55A ATOM 793 CG1 ILE A 109 27.911 17.261 8.398 1.00 35.82 A ATOM 794 CD1ILE A 109 27.901 15.782 8.661 1.00 39.05 A ATOM 795 C ILE A 109 26.42719.671 7.524 1.00 35.47 A ATOM 796 O ILE A 109 26.553 20.197 8.634 1.0034.00 A ATOM 797 N ASP A 110 25.262 19.526 6.911 1.00 35.28 A ATOM 798CA ASP A 110 24.032 20.002 7.499 1.00 36.47 A ATOM 799 CB ASP A 11023.577 21.231 6.710 1.00 39.55 A ATOM 800 CG ASP A 110 23.876 22.5267.408 1.00 42.35 A ATOM 801 OD1 ASP A 110 24.816 22.555 8.227 1.00 42.34A ATOM 802 OD2 ASP A 110 23.155 23.514 7.144 1.00 45.22 A ATOM 803 C ASPA 110 22.997 18.886 7.419 1.00 34.76 A ATOM 804 O ASP A 110 22.94418.147 6.437 1.00 31.82 A ATOM 805 N GLY A 111 22.195 18.762 8.470 1.0033.94 A ATOM 806 CA GLY A 111 21.156 17.755 8.509 1.00 35.79 A ATOM 807C GLY A 111 19.931 18.330 9.201 1.00 35.96 A ATOM 808 O GLY A 111 19.95719.465 9.672 1.00 35.30 A ATOM 809 N GLU A 112 18.855 17.553 9.244 1.0037.97 A ATOM 810 CA GLU A 112 17.613 17.962 9.893 1.00 38.66 A ATOM 811CB GLU A 112 16.502 18.123 8.850 1.00 39.80 A ATOM 812 CG GLU A 11215.336 19.019 9.264 1.00 38.95 A ATOM 813 CD GLU A 112 14.378 18.35210.240 1.00 40.97 A ATOM 814 OE1 GLU A 112 13.934 17.222 9.952 1.0041.18 A ATOM 815 OE2 GLU A 112 14.060 18.964 11.285 1.00 40.91 A ATOM816 C GLU A 112 17.311 16.810 10.845 1.00 39.79 A ATOM 817 O GLU A 11217.540 15.654 10.505 1.00 40.11 A ATOM 818 N VAL A 113 16.808 17.11812.034 1.00 41.22 A ATOM 819 CA VAL A 113 16.536 16.082 13.027 1.0042.17 A ATOM 820 CB VAL A 113 16.051 16.700 14.363 1.00 41.43 A ATOM 821CG1 VAL A 113 17.131 17.595 14.931 1.00 40.29 A ATOM 822 CG2 VAL A 11314.762 17.478 14.153 1.00 39.96 A ATOM 823 C VAL A 113 15.557 14.98912.616 1.00 43.71 A ATOM 824 O VAL A 113 15.492 13.939 13.251 1.00 45.17A ATOM 825 N THR A 114 14.802 15.222 11.553 1.00 45.21 A ATOM 826 CA THRA 114 13.843 14.226 11.104 1.00 46.88 A ATOM 827 CB THR A 114 12.41014.782 11.146 1.00 48.04 A ATOM 828 OG1 THR A 114 12.160 15.360 12.4331.00 51.16 A ATOM 829 CG2 THR A 114 11.405 13.675 10.900 1.00 50.04 AATOM 830 C THR A 114 14.126 13.764 9.681 1.00 46.51 A ATOM 831 O THR A114 13.642 12.715 9.256 1.00 47.78 A ATOM 832 N GLY A 115 14.910 14.5458.946 1.00 43.52 A ATOM 833 CA GLY A 115 15.202 14.182 7.572 1.00 43.24A ATOM 834 C GLY A 115 16.606 13.680 7.324 1.00 41.33 A ATOM 835 O GLY A115 16.873 13.058 6.294 1.00 42.99 A ATOM 836 N GLY A 116 17.507 13.9548.259 1.00 40.18 A ATOM 837 CA GLY A 116 18.881 13.509 8.102 1.00 38.86A ATOM 838 C GLY A 116 19.699 14.416 7.202 1.00 36.69 A ATOM 839 O GLY A116 19.308 15.543 6.940 1.00 36.36 A ATOM 840 N PHE A 117 20.827 13.9056.724 1.00 36.25 A ATOM 841 CA PHE A 117 21.749 14.649 5.862 1.00 35.53A ATOM 842 CB PHE A 117 22.828 13.706 5.325 1.00 34.63 A ATOM 843 CG PHEA 117 23.756 14.350 4.329 1.00 33.58 A ATOM 844 CD1 PHE A 117 24.87415.059 4.755 1.00 32.41 A ATOM 845 CD2 PHE A 117 23.502 14.255 2.9611.00 32.56 A ATOM 846 CE1 PHE A 117 25.737 15.666 3.836 1.00 31.07 AATOM 847 CE2 PHE A 117 24.357 14.859 2.032 1.00 31.84 A ATOM 848 CZ PHEA 117 25.479 15.566 2.474 1.00 31.62 A ATOM 849 C PHE A 117 21.08115.337 4.675 1.00 35.92 A ATOM 850 O PHE A 117 20.229 14.743 4.012 1.0035.06 A ATOM 851 N LYS A 118 21.491 16.576 4.395 1.00 34.72 A ATOM 852CA LYS A 118 20.937 17.315 3.265 1.00 34.75 A ATOM 853 CB LYS A 11819.691 18.098 3.698 1.00 36.47 A ATOM 854 CG LYS A 118 19.918 19.1424.783 1.00 37.72 A ATOM 855 CD LYS A 118 18.714 20.079 4.867 1.00 40.72A ATOM 856 CE LYS A 118 18.968 21.253 5.807 1.00 39.89 A ATOM 857 NZ LYSA 118 17.892 22.284 5.712 1.00 39.12 A ATOM 858 C LYS A 118 21.92018.251 2.546 1.00 32.27 A ATOM 859 O LYS A 118 21.713 18.584 1.380 1.0029.66 A ATOM 860 N LYS A 119 22.992 18.672 3.218 1.00 31.59 A ATOM 861CA LYS A 119 23.955 19.551 2.564 1.00 30.61 A ATOM 862 CB LYS A 11923.400 20.985 2.489 1.00 34.11 A ATOM 863 CG LYS A 119 22.923 21.5793.783 1.00 34.64 A ATOM 864 CD LYS A 119 22.230 22.956 3.600 1.00 33.28A ATOM 865 CE LYS A 119 23.155 23.995 2.982 1.00 32.16 A ATOM 866 NZ LYSA 119 22.817 25.404 3.328 1.00 29.97 A ATOM 867 C LYS A 119 25.38119.578 3.110 1.00 30.58 A ATOM 868 O LYS A 119 25.657 19.174 4.246 1.0028.47 A ATOM 869 N THR A 120 26.298 20.021 2.258 1.00 31.56 A ATOM 870CA THR A 120 27.702 20.156 2.637 1.00 30.27 A ATOM 871 CB THR A 12028.645 19.319 1.749 1.00 29.70 A ATOM 872 OG1 THR A 120 28.467 19.7100.381 1.00 28.77 A ATOM 873 CG2 THR A 120 28.370 17.841 1.904 1.00 28.77A ATOM 874 C THR A 120 28.071 21.605 2.417 1.00 30.38 A ATOM 875 O THR A120 27.333 22.356 1.760 1.00 29.15 A ATOM 876 N TYR A 121 29.226 21.9842.952 1.00 28.30 A ATOM 877 CA TYR A 121 29.740 23.332 2.817 1.00 29.10A ATOM 878 CB TYR A 121 29.632 24.107 4.136 1.00 30.57 A ATOM 879 CG TYRA 121 28.233 24.402 4.611 1.00 31.61 A ATOM 880 CD1 TYR A 121 27.45023.416 5.208 1.00 32.95 A ATOM 881 CE1 TYR A 121 26.160 23.692 5.6501.00 33.00 A ATOM 882 CD2 TYR A 121 27.692 25.674 4.468 1.00 33.43 AATOM 883 CE2 TYR A 121 26.406 25.959 4.903 1.00 35.44 A ATOM 884 CZ TYRA 121 25.648 24.966 5.488 1.00 33.78 A ATOM 885 OH TYR A 121 24.36925.259 5.885 1.00 36.41 A ATOM 886 C TYR A 121 31.215 23.250 2.461 1.0028.23 A ATOM 887 O TYR A 121 31.916 22.330 2.881 1.00 27.61 A ATOM 888 NALA A 122 31.675 24.212 1.679 1.00 29.43 A ATOM 889 CA ALA A 122 33.07924.284 1.324 1.00 30.31 A ATOM 890 CB ALA A 122 33.258 24.126 −0.1681.00 29.87 A ATOM 891 C ALA A 122 33.512 25.677 1.775 1.00 30.35 A ATOM892 O ALA A 122 32.971 26.675 1.305 1.00 30.95 A ATOM 893 N PHE A 12334.456 25.740 2.710 1.00 30.19 A ATOM 894 CA PHE A 123 34.945 27.0233.212 1.00 31.04 A ATOM 895 CB PHE A 123 35.185 26.982 4.728 1.00 32.77A ATOM 896 CG PHE A 123 33.937 26.889 5.543 1.00 33.65 A ATOM 897 CD1PHE A 123 33.215 25.702 5.601 1.00 34.51 A ATOM 898 CD2 PHE A 123 33.48927.988 6.273 1.00 36.59 A ATOM 899 CE1 PHE A 123 32.054 25.608 6.3841.00 36.52 A ATOM 900 CE2 PHE A 123 32.330 27.908 7.061 1.00 36.10 AATOM 901 CZ PHE A 123 31.616 26.719 7.115 1.00 34.21 A ATOM 902 C PHE A123 36.257 27.385 2.547 1.00 32.63 A ATOM 903 O PHE A 123 37.079 26.5072.265 1.00 34.75 A ATOM 904 N PHE A 124 36.467 28.676 2.312 1.00 32.86 AATOM 905 CA PHE A 124 37.706 29.123 1.702 1.00 33.91 A ATOM 906 CB PHE A124 37.410 29.930 0.441 1.00 33.32 A ATOM 907 CG PHE A 124 36.423 29.261−0.479 1.00 32.63 A ATOM 908 CD1 PHE A 124 35.106 29.712 −0.555 1.0031.58 A ATOM 909 CD2 PHE A 124 36.797 28.158 −1.241 1.00 33.35 A ATOM910 CE1 PHE A 124 34.179 29.068 −1.377 1.00 34.49 A ATOM 911 CE2 PHE A124 35.877 27.504 −2.067 1.00 33.34 A ATOM 912 CZ PHE A 124 34.56927.957 −2.135 1.00 33.17 A ATOM 913 C PHE A 124 38.495 29.959 2.703 1.0033.67 A ATOM 914 O PHE A 124 37.940 30.463 3.681 1.00 33.97 A ATOM 915 NPRO A 125 39.814 30.079 2.495 1.00 35.21 A ATOM 916 CD PRO A 125 40.64229.338 1.527 1.00 35.76 A ATOM 917 CA PRO A 125 40.660 30.866 3.397 1.0034.83 A ATOM 918 CB PRO A 125 42.048 30.718 2.776 1.00 35.56 A ATOM 919CG PRO A 125 41.995 29.336 2.207 1.00 35.73 A ATOM 920 C PRO A 12540.192 32.310 3.439 1.00 34.10 A ATOM 921 O PRO A 125 39.948 32.9252.405 1.00 34.97 A ATOM 922 N THR A 126 40.056 32.846 4.645 1.00 36.61 AATOM 923 CA THR A 126 39.605 34.217 4.822 1.00 38.10 A ATOM 924 CB THR A126 39.556 34.572 6.319 1.00 37.99 A ATOM 925 OG1 THR A 126 38.72533.620 6.991 1.00 42.36 A ATOM 926 CG2 THR A 126 38.977 35.966 6.5351.00 41.12 A ATOM 927 C THR A 126 40.492 35.218 4.085 1.00 37.53 A ATOM928 O THR A 126 40.014 36.258 3.635 1.00 37.06 A ATOM 929 N ASP A 12741.777 34.897 3.940 1.00 39.58 A ATOM 930 CA ASP A 127 42.701 35.8013.263 1.00 39.86 A ATOM 931 CB ASP A 127 44.026 35.896 4.034 1.00 42.23A ATOM 932 CG ASP A 127 44.763 34.567 4.124 1.00 43.17 A ATOM 933 OD1ASP A 127 44.243 33.539 3.641 1.00 45.25 A ATOM 934 OD2 ASP A 127 45.87834.554 4.688 1.00 45.84 A ATOM 935 C ASP A 127 42.973 35.435 1.810 1.0039.84 A ATOM 936 O ASP A 127 43.943 35.904 1.221 1.00 40.14 A ATOM 937 NASN A 128 42.109 34.600 1.238 1.00 39.79 A ATOM 938 CA ASN A 128 42.23834.182 −0.153 1.00 39.11 A ATOM 939 CB ASN A 128 43.332 33.121 −0.3051.00 40.96 A ATOM 940 CG ASN A 128 43.608 32.772 −1.763 1.00 42.76 AATOM 941 OD1 ASN A 128 44.327 31.816 −2.058 1.00 45.02 A ATOM 942 ND2ASN A 128 43.044 33.548 −2.679 1.00 43.31 A ATOM 943 C ASN A 128 40.91033.600 −0.611 1.00 38.19 A ATOM 944 O ASN A 128 40.834 32.444 −1.0271.00 36.34 A ATOM 945 N MET A 129 39.860 34.407 −0.532 1.00 37.03 A ATOM946 CA MET A 129 38.544 33.948 −0.933 1.00 36.26 A ATOM 947 CB MET A 12937.462 34.742 −0.205 1.00 35.19 A ATOM 948 CG MET A 129 37.469 34.5781.300 1.00 36.19 A ATOM 949 SD MET A 129 36.104 35.498 2.022 1.00 35.06A ATOM 950 CE MET A 129 36.869 37.133 2.125 1.00 37.35 A ATOM 951 C META 129 38.375 34.127 −2.431 1.00 35.46 A ATOM 952 O MET A 129 38.95335.036 −3.024 1.00 35.81 A ATOM 953 N PRO A 130 37.587 33.252 −3.0671.00 35.15 A ATOM 954 CD PRO A 130 37.024 31.988 −2.558 1.00 33.45 AATOM 955 CA PRO A 130 37.375 33.374 −4.510 1.00 34.52 A ATOM 956 CB PROA 130 36.935 31.968 −4.907 1.00 34.77 A ATOM 957 CG PRO A 130 36.15531.528 −3.713 1.00 33.09 A ATOM 958 C PRO A 130 36.313 34.428 −4.8251.00 35.76 A ATOM 959 O PRO A 130 35.502 34.785 −3.966 1.00 34.18 A ATOM960 N GLY A 131 36.343 34.936 −6.055 1.00 36.06 A ATOM 961 CA GLY A 13135.371 35.921 −6.483 1.00 36.41 A ATOM 962 C GLY A 131 34.313 35.190−7.282 1.00 37.83 A ATOM 963 O GLY A 131 34.424 33.981 −7.482 1.00 38.41A ATOM 964 N VAL A 132 33.288 35.900 −7.739 1.00 37.75 A ATOM 965 CA VALA 132 32.229 35.269 −8.515 1.00 39.80 A ATOM 966 CB VAL A 132 31.10636.277 −8.848 1.00 39.90 A ATOM 967 CG1 VAL A 132 30.089 35.649 −9.7921.00 39.29 A ATOM 968 CG2 VAL A 132 30.420 36.711 −7.569 1.00 39.79 AATOM 969 C VAL A 132 32.795 34.688 −9.804 1.00 40.83 A ATOM 970 O VAL A132 32.388 33.615 −10.245 1.00 41.10 A ATOM 971 N ALA A 133 33.74735.397 −10.398 1.00 42.28 A ATOM 972 CA ALA A 133 34.378 34.949 −11.6321.00 43.17 A ATOM 973 CB ALA A 133 35.482 35.923 −12.036 1.00 44.08 AATOM 974 C ALA A 133 34.955 33.548 −11.462 1.00 43.96 A ATOM 975 O ALA A133 34.636 32.651 −12.235 1.00 43.99 A ATOM 976 N GLU A 134 35.79933.366 −10.446 1.00 44.05 A ATOM 977 CA GLU A 134 36.421 32.067 −10.1731.00 44.20 A ATOM 978 CB GLU A 134 37.366 32.159 −8.969 1.00 45.02 AATOM 979 CG GLU A 134 38.507 33.150 −9.119 1.00 48.35 A ATOM 980 CD GLUA 134 38.168 34.527 −8.579 1.00 50.23 A ATOM 981 OE1 GLU A 134 37.19035.148 −9.057 1.00 51.33 A ATOM 982 OE2 GLU A 134 38.889 34.991 −7.6701.00 52.57 A ATOM 983 C GLU A 134 35.406 30.954 −9.905 1.00 42.84 A ATOM984 O GLU A 134 35.587 29.821 −10.348 1.00 44.08 A ATOM 985 N LEU A 13534.343 31.272 −9.175 1.00 41.08 A ATOM 986 CA LEU A 135 33.325 30.276−8.859 1.00 40.14 A ATOM 987 CB LEU A 135 32.349 30.827 −7.808 1.0038.00 A ATOM 988 CG LEU A 135 32.858 30.961 −6.363 1.00 35.55 A ATOM 989CD1 LEU A 135 31.813 31.634 −5.501 1.00 37.80 A ATOM 990 CD2 LEU A 13533.180 29.589 −5.807 1.00 32.97 A ATOM 991 C LEU A 135 32.544 29.786−10.079 1.00 41.44 A ATOM 992 O LEU A 135 32.419 28.583 −10.294 1.0041.03 A ATOM 993 N SER A 136 32.018 30.710 −10.877 1.00 43.65 A ATOM 994CA SER A 136 31.234 30.341 −12.064 1.00 45.88 A ATOM 995 CB SER A 13630.756 31.599 −12.798 1.00 47.35 A ATOM 996 OG SER A 136 31.851 32.390−13.227 1.00 49.44 A ATOM 997 C SER A 136 31.995 29.448 −13.041 1.0045.67 A ATOM 998 O SER A 136 31.395 28.799 −13.903 1.00 46.58 A ATOM 999N ALA A 137 33.316 29.416 −12.892 1.00 45.23 A ATOM 1000 CA ALA A 13734.185 28.618 −13.746 1.00 44.09 A ATOM 1001 CB ALA A 137 35.618 29.123−13.627 1.00 45.40 A ATOM 1002 C ALA A 137 34.128 27.127 −13.413 1.0043.23 A ATOM 1003 O ALA A 137 34.433 26.284 −14.258 1.00 42.56 A ATOM1004 N ILE A 138 33.758 26.799 −12.179 1.00 41.63 A ATOM 1005 CA ILE A138 33.664 25.398 −11.775 1.00 40.86 A ATOM 1006 CB ILE A 138 33.26025.275 −10.280 1.00 39.46 A ATOM 1007 CG2 ILE A 138 33.120 23.812 −9.8931.00 39.81 A ATOM 1008 CG1 ILE A 138 34.319 25.955 −9.403 1.00 40.37 AATOM 1009 CD1 ILE A 138 33.990 25.985 −7.921 1.00 38.95 A ATOM 1010 CILE A 138 32.600 24.771 −12.677 1.00 40.15 A ATOM 1011 O ILE A 13831.481 25.261 −12.745 1.00 39.48 A ATOM 1012 N PRO A 139 32.949 23.688−13.391 1.00 42.04 A ATOM 1013 CD PRO A 139 34.277 23.056 −13.319 1.0042.63 A ATOM 1014 CA PRO A 139 32.080 22.950 −14.325 1.00 42.17 A ATOM1015 CB PRO A 139 32.913 21.716 −14.668 1.00 43.33 A ATOM 1016 CG PRO A139 34.308 22.234 −14.591 1.00 44.53 A ATOM 1017 C PRO A 139 30.67222.568 −13.873 1.00 41.62 A ATOM 1018 O PRO A 139 29.717 22.686 −14.6461.00 41.11 A ATOM 1019 N SER A 140 30.548 22.103 −12.634 1.00 40.71 AATOM 1020 CA SER A 140 29.262 21.677 −12.095 1.00 40.02 A ATOM 1021 CBSER A 140 29.458 20.440 −11.207 1.00 38.55 A ATOM 1022 OG SER A 14030.473 20.655 −10.239 1.00 37.26 A ATOM 1023 C SER A 140 28.510 22.765−11.329 1.00 40.37 A ATOM 1024 O SER A 140 27.460 22.507 −10.735 1.0040.00 A ATOM 1025 N MET A 141 29.044 23.981 −11.351 1.00 39.35 A ATOM1026 CA MET A 141 28.409 25.107 −10.671 1.00 38.79 A ATOM 1027 CB MET A141 29.383 26.292 −10.608 1.00 38.90 A ATOM 1028 CG MET A 141 28.90427.492 −9.790 1.00 39.10 A ATOM 1029 SD MET A 141 28.779 27.156 −8.0061.00 38.89 A ATOM 1030 CE MET A 141 30.511 27.219 −7.524 1.00 39.82 AATOM 1031 C MET A 141 27.167 25.500 −11.474 1.00 38.43 A ATOM 1032 O META 141 27.168 25.418 −12.704 1.00 38.34 A ATOM 1033 N PRO A 142 26.08825.924 −10.794 1.00 38.99 A ATOM 1034 CD PRO A 142 25.876 26.042 −9.3401.00 37.35 A ATOM 1035 CA PRO A 142 24.882 26.314 −11.535 1.00 37.96 AATOM 1036 CB PRO A 142 23.949 26.823 −10.439 1.00 38.94 A ATOM 1037 CGPRO A 142 24.372 26.043 −9.235 1.00 37.01 A ATOM 1038 C PRO A 142 25.21027.413 −12.545 1.00 39.39 A ATOM 1039 O PRO A 142 25.983 28.329 −12.2481.00 38.95 A ATOM 1040 N PRO A 143 26.644 27.328 −13.758 1.00 37.81 AATOM 1041 CD PRO A 143 23.843 26.222 −14.307 1.00 39.42 A ATOM 1042 CAPRO A 143 24.903 28.344 −14.780 1.00 38.06 A ATOM 1043 CB PRO A 14324.060 27.863 −15.964 1.00 38.66 A ATOM 1044 CG PRO A 143 24.053 26.383−15.793 1.00 39.51 A ATOM 1045 C PRO A 143 24.439 29.708 −14.266 1.0038.30 A ATOM 1046 O PRO A 143 24.955 30.755 −14.665 1.00 37.08 A ATOM1047 N ALA A 144 23.462 29.670 −13.364 1.00 37.27 A ATOM 1048 CA ALA A144 22.887 30.871 −12.773 1.00 37.43 A ATOM 1049 CB ALA A 144 21.81330.485 −11.767 1.00 35.79 A ATOM 1050 C ALA A 144 23.910 31.793 −12.1151.00 36.34 A ATOM 1051 O ALA A 144 23.700 33.002 −12.047 1.00 37.01 AATOM 1052 N VAL A 145 25.018 31.241 −11.636 1.00 35.52 A ATOM 1053 CAVAL A 145 26.020 32.087 −10.993 1.00 36.56 A ATOM 1054 CB VAL A 14527.105 31.231 −10.292 1.00 33.87 A ATOM 1055 CG1 VAL A 145 28.206 32.118−9.755 1.00 32.14 A ATOM 1056 CG2 VAL A 145 26.475 30.421 −9.164 1.0033.34 A ATOM 1057 C VAL A 145 26.674 33.047 −12.000 1.00 37.24 A ATOM1058 O VAL A 145 26.787 34.246 −11.746 1.00 35.40 A ATOM 1059 N ALA A146 27.093 32.524 −13.146 1.00 38.95 A ATOM 1060 CA ALA A 146 27.71333.365 −14.165 1.00 40.89 A ATOM 1061 CB ALA A 146 28.233 32.505 −15.3231.00 41.08 A ATOM 1062 C ALA A 146 26.695 34.374 −14.679 1.00 41.46 AATOM 1063 O ALA A 146 27.013 35.543 −14.889 1.00 41.33 A ATOM 1064 N GLUA 147 25.464 33.917 −14.866 1.00 42.76 A ATOM 1065 CA GLU A 147 24.40534.777 −15.367 1.00 44.04 A ATOM 1066 CB GLU A 147 23.161 33.938 −15.6571.00 46.32 A ATOM 1067 CG GLU A 147 23.296 33.089 −16.924 1.00 51.66 AATOM 1068 CD GLU A 147 22.729 31.687 −16.774 1.00 53.70 A ATOM 1069 OE1GLU A 147 21.566 31.555 −16.334 1.00 55.88 A ATOM 1070 OE2 GLU A 14723.449 30.717 −17.104 1.00 56.34 A ATOM 1071 C GLU A 147 24.066 35.923−14.421 1.00 43.26 A ATOM 1072 O GLU A 147 23.458 36.910 −14.833 1.0041.77 A ATOM 1073 N ASN A 148 24.474 35.797 −13.162 1.00 42.18 A ATOM1074 CA ASN A 148 24.200 36.827 −12.165 1.00 41.99 A ATOM 1075 CB ASN A148 23.588 36.193 −10.911 1.00 42.15 A ATOM 1076 CG ASN A 148 22.10435.918 −11.059 1.00 40.63 A ATOM 1077 OD1 ASN A 148 21.301 36.841−11.145 1.00 42.33 A ATOM 1078 ND2 ASN A 148 21.735 34.649 −11.093 1.0041.13 A ATOM 1079 C ASN A 148 25.435 37.630 −11.774 1.00 41.70 A ATOM1080 O ASN A 148 25.361 38.492 −10.904 1.00 41.52 A ATOM 1081 N ALA A149 26.560 37.350 −12.427 1.00 42.05 A ATOM 1082 CA ALA A 149 27.82538.027 −12.150 1.00 42.82 A ATOM 1083 CB ALA A 149 28.898 37.556 −13.1431.00 41.85 A ATOM 1084 C ALA A 149 27.743 39.553 −12.169 1.00 43.54 AATOM 1085 O ALA A 149 28.236 40.217 −11.257 1.00 44.06 A ATOM 1086 N GLUA 150 27.132 40.103 −13.215 1.00 44.98 A ATOM 1087 CA GLU A 150 26.99741.549 −13.355 1.00 45.33 A ATOM 1088 CB GLU A 150 26.373 41.891 −14.7081.00 47.44 A ATOM 1089 CG GLU A 150 27.297 41.639 −15.876 1.00 52.19 AATOM 1090 CD GLU A 150 26.605 41.834 −17.206 1.00 54.21 A ATOM 1091 OE1GLU A 150 25.720 41.018 −17.540 1.00 56.56 A ATOM 1092 OE2 GLU A 15026.936 42.808 −17.912 1.00 54.57 A ATOM 1093 C GLU A 150 26.156 42.145−12.241 1.00 43.75 A ATOM 1094 O GLU A 150 26.485 43.200 −11.695 1.0043.51 A ATOM 1095 N LEU A 151 25.063 41.468 −11.915 1.00 42.63 A ATOM1096 CA LEU A 151 24.181 41.922 −10.853 1.00 41.40 A ATOM 1097 CB LEU A151 22.963 40.995 −10.777 1.00 42.33 A ATOM 1098 CG LEU A 151 21.80441.368 −9.850 1.00 42.54 A ATOM 1099 CD1 LEU A 151 20.538 40.668 −10.3231.00 43.25 A ATOM 1100 CD2 LEU A 151 22.139 40.989 −8.414 1.00 41.89 AATOM 1101 C LEU A 151 24.954 41.941 −9.522 1.00 40.27 A ATOM 1102 O LEUA 151 24.973 42.956 −8.822 1.00 41.45 A ATOM 1103 N PHE A 152 25.59740.826 −9.181 1.00 38.34 A ATOM 1104 CA PHE A 152 26.372 40.745 −7.9461.00 36.94 A ATOM 1105 CB PHE A 152 27.129 39.417 −7.870 1.00 34.62 AATOM 1106 CG PHE A 152 26.249 38.217 −7.670 1.00 33.59 A ATOM 1107 CD1PHE A 152 26.625 36.981 −8.183 1.00 32.62 A ATOM 1108 CD2 PHE A 15225.071 38.307 −6.936 1.00 31.47 A ATOM 1109 CE1 PHE A 152 25.842 35.844−7.969 1.00 32.66 A ATOM 1110 CE2 PHE A 152 24.281 37.177 −6.714 1.0031.36 A ATOM 1111 CZ PHE A 152 24.670 35.941 −7.232 1.00 32.67 A ATOM1112 C PHE A 152 27.379 41.891 −7.889 1.00 38.16 A ATOM 1113 O PHE A 15227.499 42.575 −6.874 1.00 38.05 A ATOM 1114 N ALA A 153 28.098 42.094−8.989 1.00 38.70 A ATOM 1115 CA ALA A 153 29.106 43.145 −9.066 1.0039.88 A ATOM 1116 CB ALA A 153 29.783 43.117 −10.433 1.00 41.69 A ATOM1117 C ALA A 153 28.523 44.526 −8.805 1.00 40.13 A ATOM 1118 O ALA A 15329.120 45.342 −8.101 1.00 40.16 A ATOM 1119 N ARG A 154 27.350 44.777−9.374 1.00 40.05 A ATOM 1120 CA ARG A 154 26.682 46.055 −9.217 1.0040.63 A ATOM 1121 CB ARG A 154 25.394 46.057 −10.044 1.00 42.51 A ATOM1122 CG ARG A 154 24.770 47.425 −10.221 1.00 47.49 A ATOM 1123 CD ARG A154 24.025 47.541 −11.555 1.00 48.14 A ATOM 1124 NE ARG A 154 23.06046.463 −11.764 1.00 48.78 A ATOM 1125 CZ ARG A 154 23.298 45.368 −12.4821.00 46.90 A ATOM 1126 NH1 ARG A 154 24.473 45.197 −13.069 1.00 48.01 AATOM 1127 NH2 ARG A 154 22.359 44.443 −12.617 1.00 46.15 A ATOM 1128 CARG A 154 26.387 46.397 −7.750 1.00 40.64 A ATOM 1129 O ARG A 154 26.29747.574 −7.387 1.00 40.09 A ATOM 1130 N TYR A 155 26.256 45.383 −6.8971.00 4035 A ATOM 1131 CA TYR A 155 25.975 45.652 −5.493 1.00 38.19 AATOM 1132 CB TYR A 155 24.720 44.901 −5.052 1.00 38.67 A ATOM 1133 CGTYR A 155 23.503 45.381 −5.790 1.00 38.24 A ATOM 1134 CD1 TYR A 15523.171 44.855 −7.035 1.00 38.78 A ATOM 1135 CE1 TYR A 155 22.099 45.357−7.766 1.00 39.06 A ATOM 1136 CD2 TYR A 155 22.725 46.424 −5.284 1.0038.96 A ATOM 1137 CE2 TYR A 155 21.651 46.935 −6.006 1.00 40.27 A ATOM1138 CZ TYR A 155 21.346 46.398 −7.249 1.00 40.64 A ATOM 1139 OH TYR A155 20.301 46.910 −7.983 1.00 42.74 A ATOM 1140 C TYR A 155 27.12945.367 −4.545 1.00 38.20 A ATOM 1141 O TYR A 155 26.957 45.356 −3.3251.00 38.12 A ATOM 1142 N GLY A 156 28.311 45.147 −5.105 1.00 37.37 AATOM 1143 CA GLY A 156 29.471 44.904 −4.270 1.00 37.64 A ATOM 1144 C GLYA 156 29.602 43.510 −3.700 1.00 37.28 A ATOM 1145 O GLY A 156 30.44943.274 −2.835 1.00 37.51 A ATOM 1146 N LEU A 157 28.765 42.586 −4.1571.00 36.82 A ATOM 1147 CA LEU A 157 28.846 41.210 −3.682 1.00 36.96 AATOM 1148 CB LEU A 157 27.531 40.480 −3.947 1.00 35.41 A ATOM 1149 CGLEU A 157 26.314 41.101 −3.245 1.00 35.60 A ATOM 1150 CD1 LEU A 15725.045 40.359 −3.646 1.00 33.80 A ATOM 1151 CD2 LEU A 157 26.508 41.048−1.741 1.00 34.21 A ATOM 1152 C LEU A 157 29.984 40.588 −4.479 1.0037.62 A ATOM 1153 O LEU A 157 29.814 40.233 −5.641 1.00 38.02 A ATOM1154 N ASP A 158 31.147 40.466 −3.847 1.00 37.12 A ATOM 1155 CA ASP A158 32.323 39.942 −4.526 1.00 37.41 A ATOM 1156 CB ASP A 158 33.35541.083 −4.669 1.00 38.84 A ATOM 1157 CG ASP A 158 34.737 40.598 −5.1061.00 39.06 A ATOM 1158 OD1 ASP A 158 34.823 39.688 −5.952 1.00 40.99 AATOM 1159 OD2 ASP A 158 35.743 41.150 −4.610 1.00 42.10 A ATOM 1160 CASP A 158 32.948 38.713 −3.871 1.00 37.01 A ATOM 1161 O ASP A 158 32.82037.600 −4.386 1.00 37.39 A ATOM 1162 N LYS A 159 33.608 38.902 −2.7341.00 35.40 A ATOM 1163 CA LYS A 159 34.258 37.790 −2.061 1.00 34.29 AATOM 1164 CB LYS A 159 35.313 38.316 −1.083 1.00 35.64 A ATOM 1165 CGLYS A 159 36.464 39.047 −1.782 1.00 35.48 A ATOM 1166 CD LYS A 15937.021 38.202 −2.931 1.00 39.05 A ATOM 1167 CE LYS A 159 38.115 38.924−3.701 1.00 42.30 A ATOM 1168 NZ LYS A 159 37.950 38.741 −5.171 1.0044.17 A ATOM 1169 C LYS A 159 33.312 36.824 −1.362 1.00 33.08 A ATOM1170 O LYS A 159 32.411 37.218 −0.615 1.00 32.50 A ATOM 1171 N VAL A 16033.555 35.546 −1.615 1.00 33.33 A ATOM 1172 CA VAL A 160 32.764 34.453−1.071 1.00 33.25 A ATOM 1173 CB VAL A 160 32.317 33.542 −2.223 1.0032.90 A ATOM 1174 CG1 VAL A 160 31.591 32.306 −1.698 1.00 33.69 A ATOM1175 CG2 VAL A 160 31.429 34.344 −3.162 1.00 30.05 A ATOM 1176 C VAL A160 33.574 33.668 −0.045 1.00 33.60 A ATOM 1177 O VAL A 160 34.60833.086 −0.375 1.00 33.21 A ATOM 1178 N GLN A 161 33.105 33.681 1.2041.00 33.10 A ATOM 1179 CA GLN A 161 33.760 32.981 2.308 1.00 34.17 AATOM 1180 CB GLN A 161 33.308 33.587 3.645 1.00 33.46 A ATOM 1181 CG GLNA 161 33.870 32.894 4.882 1.00 38.50 A ATOM 1182 CD GLN A 161 35.38433.046 5.024 1.00 38.34 A ATOM 1183 OE1 GLN A 161 35.883 34.116 5.3651.00 42.77 A ATOM 1184 NE2 GLN A 161 36.115 31.972 4.753 1.00 40.54 AATOM 1185 C GLN A 161 33.479 31.470 2.297 1.00 33.79 A ATOM 1186 O GLN A161 34.224 30.684 2.893 1.00 33.36 A ATOM 1187 N MET A 162 32.401 31.0651.629 1.00 31.86 A ATOM 1188 CA MET A 162 32.056 29.649 1.536 1.00 30.83A ATOM 1189 CB MET A 162 31.812 29.048 2.925 1.00 34.42 A ATOM 1190 CGMET A 162 30.424 29.337 3.461 1.00 36.50 A ATOM 1191 SD MET A 162 30.43330.247 5.003 1.00 43.69 A ATOM 1192 CE MET A 162 29.357 29.180 5.9781.00 40.85 A ATOM 1193 C MET A 162 30.804 29.439 0.694 1.00 28.08 A ATOM1194 O MET A 162 30.068 30.373 0.416 1.00 29.84 A ATOM 1195 N THR A 16330.585 28.200 0.278 1.00 27.90 A ATOM 1196 CA THR A 163 29.404 27.862−0.501 1.00 28.23 A ATOM 1197 CB THR A 163 29.753 27.471 −1.969 1.0028.07 A ATOM 1198 OG1 THR A 163 30.534 26.268 −1.981 1.00 28.96 A ATOM1199 CG2 THR A 163 30.515 28.579 −2.640 1.00 28.00 A ATOM 1200 C THR A163 28.781 26.657 0.160 1.00 28.16 A ATOM 1201 O THR A 163 29.440 25.9600.938 1.00 27.88 A ATOM 1202 N SER A 164 27.500 26.426 −0.117 1.00 27.41A ATOM 1203 CA SER A 164 26.840 25.255 0.412 1.00 28.60 A ATOM 1204 CBSER A 164 25.893 25.594 1.574 1.00 27.04 A ATOM 1205 OG SER A 164 24.74726.310 1.159 1.00 26.10 A ATOM 1206 C SER A 164 26.078 24.620 −0.7351.00 28.66 A ATOM 1207 O SER A 164 25.693 25.301 −1.693 1.00 27.23 AATOM 1208 N MET A 165 25.909 23.306 −0.630 1.00 29.00 A ATOM 1209 CA META 165 25.199 22.495 −1.606 1.00 31.67 A ATOM 1210 CB MET A 165 26.15721.497 −2.263 1.00 31.92 A ATOM 1211 CG MET A 165 26.990 22.068 −3.3961.00 35.54 A ATOM 1212 SD MET A 165 28.324 23.135 −2.843 1.00 41.52 AATOM 1213 CE MET A 165 29.438 21.940 −2.325 1.00 31.41 A ATOM 1214 C META 165 24.075 21.727 −0.920 1.00 29.72 A ATOM 1215 O MET A 165 24.31520.961 0.013 1.00 27.76 A ATOM 1216 N ASP A 166 22.844 21.947 −1.3751.00 30.14 A ATOM 1217 CA ASP A 166 21.698 21.252 −0.814 1.00 29.25 AATOM 1218 CB ASP A 166 20.522 22.221 −0.637 1.00 31.09 A ATOM 1219 CGASP A 166 19.320 21.570 0.030 1.00 28.72 A ATOM 1220 OD1 ASP A 16619.106 20.354 −0.163 1.00 28.28 A ATOM 1221 OD2 ASP A 166 18.580 22.2840.733 1.00 30.52 A ATOM 1222 C ASP A 166 21.359 20.167 −1.826 1.00 28.71A ATOM 1223 O ASP A 166 20.857 20.458 −2.918 1.00 27.49 A ATOM 1224 NTYR A 167 21.650 18.923 −1.457 1.00 29.03 A ATOM 1225 CA TYR A 16721.433 17.766 −2.319 1.00 30.30 A ATOM 1226 CB TYR A 167 22.224 16.577−1.777 1.00 30.78 A ATOM 1227 CG TYR A 167 23.710 16.837 −1.744 1.0030.91 A ATOM 1228 CD1 TYR A 167 24.500 16.645 −2.889 1.00 29.35 A ATOM1229 CE1 TYR A 167 25.854 16.960 −2.888 1.00 30.22 A ATOM 1230 CD2 TYR A167 24.320 17.351 −0.597 1.00 28.48 A ATOM 1231 CE2 TYR A 167 25.67617.672 −0.591 1.00 29.07 A ATOM 1232 CZ TYR A 167 26.436 17.475 −1.7351.00 28.34 A ATOM 1233 OH TYR A 167 27.774 17.779 −1.732 1.00 28.84 AATOM 1234 C TYR A 167 19.974 17.371 −2.530 1.00 32.95 A ATOM 1235 O TYRA 167 19.651 16.750 −3.531 1.00 31.97 A ATOM 1236 N LYS A 168 19.09617.730 −1.596 1.00 32.49 A ATOM 1237 CA LYS A 168 17.678 17.408 −1.7311.00 34.77 A ATOM 1238 CB LYS A 168 17.017 17.311 −0.355 1.00 34.50 AATOM 1239 CG LYS A 168 17.505 16.127 0.448 1.00 35.07 A ATOM 1240 CD LYSA 168 16.738 15.999 1.750 1.00 38.33 A ATOM 1241 CE LYS A 168 17.05214.688 2.438 1.00 39.81 A ATOM 1242 NZ LYS A 168 16.297 14.561 3.7141.00 41.97 A ATOM 1243 C LYS A 168 16.937 18.427 −2.588 1.00 33.66 AATOM 1244 O LYS A 168 16.064 18.070 −3.368 1.00 35.97 A ATOM 1245 N LYSA 169 17.285 19.696 −2.451 1.00 33.56 A ATOM 1246 CA LYS A 169 16.62320.720 −3.231 1.00 33.68 A ATOM 1247 CB LYS A 169 16.364 21.946 −2.3571.00 34.85 A ATOM 1248 CG LYS A 169 15.481 21.625 −1.168 1.00 39.08 AATOM 1249 CD LYS A 169 15.060 22.866 −0.415 1.00 42.71 A ATOM 1250 CELYS A 169 14.149 22.503 0.749 1.00 44.29 A ATOM 1251 NZ LYS A 169 13.66623.718 1.473 1.00 47.87 A ATOM 1252 C LYS A 169 17.424 21.113 −4.4591.00 32.16 A ATOM 1253 O LYS A 169 16.948 21.880 −5.291 1.00 32.91 AATOM 1254 N ARG A 170 18.635 20.573 −4.573 1.00 30.70 A ATOM 1255 CA ARGA 170 19.519 20.889 −5.691 1.00 31.30 A ATOM 1256 CB ARG A 170 18.95920.322 −6.999 1.00 34.37 A ATOM 1257 CG ARG A 170 18.911 18.794 −7.0481.00 40.86 A ATOM 1258 CD ARG A 170 18.230 18.309 −8.329 1.00 45.92 AATOM 1259 NE ARG A 170 18.162 16.848 −8.360 1.00 51.57 A ATOM 1260 CZARG A 170 18.496 16.048 −7.350 1.00 53.83 A ATOM 1261 NH1 ARG A 17018.930 16.558 −6.207 1.00 55.24 A ATOM 1262 NH2 ARG A 170 18.394 14.733−7.489 1.00 55.41 A ATOM 1263 C ARG A 170 19.684 22.403 −5.793 1.0030.20 A ATOM 1264 O ARG A 170 19.392 23.012 −6.827 1.00 27.37 A ATOM1265 N GLN A 171 20.157 23.006 −4.702 1.00 27.02 A ATOM 1266 CA GLN A171 20.361 24.446 −4.634 1.00 27.26 A ATOM 1267 CB GLN A 171 19.27125.112 −3.784 1.00 27.34 A ATOM 1268 CG GLN A 171 17.861 24.995 −4.3731.00 29.77 A ATOM 1269 CD GLN A 171 16.795 25.663 −3.519 1.00 31.84 AATOM 1270 OE1 GLN A 171 16.885 25.688 −2.293 1.00 30.82 A ATOM 1271 NE2GLN A 171 15.762 26.192 −4.171 1.00 33.68 A ATOM 1272 C GLN A 171 21.73124.751 −4.044 1.00 26.95 A ATOM 1273 O GLN A 171 22.286 23.940 −3.3031.00 24.99 A ATOM 1274 N VAL A 172 22.264 25.918 −4.397 1.00 26.41 AATOM 1275 CA VAL A 172 23.573 26.369 −3.936 1.00 29.58 A ATOM 1276 CBVAL A 172 24.593 26.436 −5.114 1.00 27.24 A ATOM 1277 CG1 VAL A 17225.931 26.995 −4.627 1.00 28.16 A ATOM 1278 CG2 VAL A 172 24.802 25.061−5.703 1.00 27.97 A ATOM 1279 C VAL A 172 23.476 27.761 −3.309 1.0030.14 A ATOM 1280 O VAL A 172 22.788 28.645 −3.836 1.00 32.05 A ATOM1281 N ASN A 173 24.153 27.941 −2.176 1.00 29.48 A ATOM 1282 CA ASN A173 24.182 29.228 −1.490 1.00 27.27 A ATOM 1283 CB ASN A 173 23.88729.074 0.014 1.00 29.16 A ATOM 1284 CG ASN A 173 22.423 28.831 0.3081.00 28.68 A ATOM 1285 OD1 ASN A 173 21.594 28.860 −0.595 1.00 27.97 AATOM 1286 ND2 ASN A 173 22.094 28.599 1.591 1.00 26.44 A ATOM 1287 C ASNA 173 25.588 29.809 −1.639 1.00 27.49 A ATOM 1288 O ASN A 173 26.57229.080 −1.523 1.00 27.57 A ATOM 1289 N LEU A 174 25.672 31.109 −1.9071.00 26.38 A ATOM 1290 CA LEU A 174 26.961 31.793 −2.011 1.00 27.88 AATOM 1291 CB LEU A 174 27.047 32.605 −3.308 1.00 29.77 A ATOM 1292 CGLEU A 174 26.831 31.854 −4.631 1.00 29.34 A ATOM 1293 CD1 LEU A 17427.115 32.800 −5.784 1.00 30.72 A ATOM 1294 CD2 LEU A 174 27.750 30.647−4.718 1.00 29.99 A ATOM 1295 C LEU A 174 27.041 32.735 −0.810 1.0027.33 A ATOM 1296 O LEU A 174 26.324 33.737 −0.749 1.00 24.91 A ATOM1297 N TYR A 175 27.906 32.420 0.151 1.00 27.35 A ATOM 1298 CA TYR A 17528.023 33.266 1.330 1.00 28.53 A ATOM 1299 CB TYR A 175 28.377 32.4202.565 1.00 26.66 A ATOM 1300 CG TYR A 175 27.343 31.347 2.890 1.00 28.27A ATOM 1301 CD1 TYR A 175 27.339 30.115 2.229 1.00 28.80 A ATOM 1302 CE1TYR A 175 26.352 29.150 2.496 1.00 29.04 A ATOM 1303 CD2 TYR A 17526.340 31.588 3.826 1.00 29.47 A ATOM 1304 CE2 TYR A 175 25.355 30.6424.094 1.00 29.90 A ATOM 1305 CZ TYR A 175 25.360 29.432 3.433 1.00 30.82A ATOM 1306 OH TYR A 175 24.358 28.522 3.702 1.00 28.60 A ATOM 1307 CTYR A 175 29.043 34.380 1.109 1.00 28.09 A ATOM 1308 O TYR A 175 30.23934.182 1.298 1.00 31.10 A ATOM 1309 N PHE A 176 28.550 35.551 0.704 1.0027.31 A ATOM 1310 CA PHE A 176 29.393 36.714 0.444 1.00 28.26 A ATOM1311 CB PHE A 176 28.622 37.774 −0.341 1.00 27.28 A ATOM 1312 CG PHE A176 28.354 37.387 −1.772 1.00 27.93 A ATOM 1313 CD1 PHE A 176 27.08136.992 −2.173 1.00 28.47 A ATOM 1314 CD2 PHE A 176 29.373 37.435 −2.7191.00 28.88 A ATOM 1315 CE1 PHE A 176 26.822 36.650 −3.511 1.00 29.48 AATOM 1316 CE2 PHE A 176 29.126 37.096 −4.062 1.00 28.83 A ATOM 1317 CZPHE A 176 27.848 36.704 −4.452 1.00 28.88 A ATOM 1318 C PHE A 176 29.91637.315 1.744 1.00 29.45 A ATOM 1319 O PHE A 176 29.180 37.418 2.734 1.0027.17 A ATOM 1320 N SER A 177 31.184 37.726 1.722 1.00 30.00 A ATOM 1321CA SER A 177 31.841 38.281 2.910 1.00 31.18 A ATOM 1322 CB SER A 17732.876 37.270 3.419 1.00 30.63 A ATOM 1323 OG SER A 177 33.179 37.4724.786 1.00 31.80 A ATOM 1324 C SER A 177 32.516 39.631 2.637 1.00 32.16A ATOM 1325 O SER A 177 32.466 40.149 1.513 1.00 31.15 A ATOM 1326 N GLUA 178 33.161 40.188 3.663 1.00 32.64 A ATOM 1327 CA GLU A 178 33.82241.485 3.542 1.00 33.04 A ATOM 1328 CB GLU A 178 35.134 41.352 2.7431.00 35.78 A ATOM 1329 CG GLU A 178 36.116 40.349 3.361 1.00 36.71 AATOM 1330 CD GLU A 178 37.519 40.402 2.759 1.00 37.67 A ATOM 1331 OE1GLU A 178 37.654 40.565 1.521 1.00 36.25 A ATOM 1332 OE2 GLU A 17838.490 40.259 3.535 1.00 37.28 A ATOM 1333 C GLU A 178 32.866 42.4582.856 1.00 33.20 A ATOM 1334 O GLU A 178 33.208 43.106 1.860 1.00 65.37A ATOM 1335 N LEU A 179 31.653 42.548 3.395 1.00 32.06 A ATOM 1336 CALEU A 179 30.623 43.422 2.843 1.00 32.29 A ATOM 1337 CB LEU A 179 29.26143.035 3.417 1.00 31.59 A ATOM 1338 CG LEU A 179 28.898 41.550 3.3031.00 32.74 A ATOM 1339 CD1 LEU A 179 27.623 41.288 4.080 1.00 34.49 AATOM 1340 CD2 LEU A 179 28.736 41.160 1.826 1.00 32.26 A ATOM 1341 C LEUA 179 30.889 44.900 3.128 1.00 33.85 A ATOM 1342 O LEU A 179 31.07245.295 4.282 1.00 33.79 A ATOM 1343 N SER A 180 30.905 45.717 2.078 1.0034.71 A ATOM 1344 CA SER A 180 31.132 47.149 2.240 1.00 36.87 A ATOM1345 CB SER A 180 31.390 47.821 0.890 1.00 38.51 A ATOM 1346 OG SER A180 30.177 48.062 0.205 1.00 41.84 A ATOM 1347 C SER A 180 29.888 47.7702.864 1.00 38.21 A ATOM 1348 O SER A 180 28.874 47.236 2.723 1.00 35.22A ATOM 1349 N ALA A 181 30.073 48.898 3.544 1.00 37.72 A ATOM 1350 CAALA A 181 28.969 49.597 4.188 1.00 39.03 A ATOM 1351 CB ALA A 181 29.48750.855 4.886 1.00 37.87 A ATOM 1352 C ALA A 181 27.901 49.968 3.166 1.0039.40 A ATOM 1353 O ALA A 181 26.701 49.909 3.449 1.00 39.97 A ATOM 1354N GLN A 182 28.342 50.337 1.971 1.00 38.70 A ATOM 1355 CA GLN A 18227.419 50.733 0.918 1.00 39.60 A ATOM 1356 CB GLN A 182 28.192 51.136−0.336 1.00 40.99 A ATOM 1357 CG GLN A 182 27.303 51.450 −1.522 1.0045.59 A ATOM 1358 CD GLN A 182 27.915 52.474 −2.455 1.00 46.88 A ATOM1359 OE1 GLN A 182 29.124 52.485 −2.683 1.00 51.04 A ATOM 1360 NE2 GLN A182 27.075 53.336 −3.013 1.00 49.22 A ATOM 1361 C GLN A 182 26.43049.634 0.580 1.00 39.40 A ATOM 1362 O GLN A 182 25.218 49.869 0.548 1.0040.56 A ATOM 1363 N THR A 183 26.951 48.435 0.330 1.00 38.08 A ATOM 1364CA THR A 183 26.125 47.284 −0.006 1.00 36.74 A ATOM 1365 CB THR A 18326.980 45.990 −0.097 1.00 36.40 A ATOM 1366 OG1 THR A 183 27.950 46.122−1.146 1.00 36.47 A ATOM 1367 CG2 THR A 183 26.098 44.789 −0.378 1.0034.06 A ATOM 1368 C THR A 183 25.003 47.045 1.005 1.00 36.17 A ATOM 1369O THR A 183 23.900 46.661 0.627 1.00 35.97 A ATOM 1370 N LEU A 18425.282 47.278 2.284 1.00 35.23 A ATOM 1371 CA LEU A 184 24.287 47.0513.327 1.00 35.08 A ATOM 1372 CB LEU A 184 24.969 46.532 4.598 1.00 34.37A ATOM 1373 CG LEU A 184 25.642 45.161 4.475 1.00 32.38 A ATOM 1374 CD1LEU A 184 26.367 44.840 5.767 1.00 34.20 A ATOM 1375 CD2 LEU A 18424.609 44.089 4.153 1.00 31.86 A ATOM 1376 C LEU A 184 23.397 48.2413.681 1.00 37.71 A ATOM 1377 O LEU A 184 22.552 48.128 4.574 1.00 36.83A ATOM 1378 N GLU A 185 23.577 49.375 3.007 1.00 37.02 A ATOM 1379 CAGLU A 185 22.726 50.525 3.296 1.00 39.60 A ATOM 1380 CB GLU A 185 23.32551.831 2.771 1.00 42.09 A ATOM 1381 CG GLU A 185 24.105 52.608 3.8191.00 47.09 A ATOM 1382 CD GLU A 185 23.375 52.699 5.158 1.00 49.21 AATOM 1383 OE1 GLU A 185 22.140 52.931 5.168 1.00 50.57 A ATOM 1384 OE2GLU A 185 24.040 52.546 6.206 1.00 48.86 A ATOM 1385 C GLU A 185 21.36950.296 2.660 1.00 37.48 A ATOM 1386 O GLU A 185 21.261 49.592 1.656 1.0035.55 A ATOM 1387 N ALA A 186 20.342 50.898 3.257 1.00 37.32 A ATOM 1388CA ALA A 186 18.965 50.754 2.795 1.00 35.80 A ATOM 1389 CB ALA A 18618.073 51.738 3.535 1.00 35.92 A ATOM 1390 C ALA A 186 18.777 50.9151.290 1.00 36.43 A ATOM 1391 O ALA A 186 18.143 50.080 0.645 1.00 34.72A ATOM 1392 N GLU A 187 19.316 51.995 0.734 1.00 37.69 A ATOM 1393 CAGLU A 187 19.183 52.254 −0.690 1.00 38.57 A ATOM 1394 CB GLU A 18719.920 53.549 −1.042 1.00 42.29 A ATOM 1395 CG GLU A 187 20.121 53.794−2.529 1.00 46.35 A ATOM 1396 CD GLU A 187 20.874 55.089 −2.798 1.0048.77 A ATOM 1397 OE1 GLU A 187 20.265 56.173 −2.642 1.00 51.07 A ATOM1398 OE2 GLU A 187 22.075 55.019 −3.149 1.00 49.51 A ATOM 1399 C GLU A187 19.697 51.092 −1.536 1.00 37.10 A ATOM 1400 O GLU A 187 19.03650.672 −2.482 1.00 36.07 A ATOM 1401 N SER A 188 20.874 50.580 −1.1951.00 36.10 A ATOM 1402 CA SER A 188 21.476 49.461 −1.914 1.00 34.79 AATOM 1403 CB SER A 188 22.916 49.240 −1.439 1.00 35.13 A ATOM 1404 OGSER A 188 23.412 47.986 −1.888 1.00 42.19 A ATOM 1405 C SER A 188 20.68548.161 −1.749 1.00 33.67 A ATOM 1406 O SER A 188 20.430 47.450 −2.7251.00 32.43 A ATOM 1407 N VAL A 189 20.308 47.846 −0.512 1.00 32.53 AATOM 1408 CA VAL A 189 19.548 46.633 −0.241 1.00 33.25 A ATOM 1409 CBVAL A 189 19.229 46.498 1.269 1.00 34.36 A ATOM 1410 CG1 VAL A 18918.267 45.345 1.500 1.00 32.73 A ATOM 1411 CG2 VAL A 189 20.512 46.2592.052 1.00 35.61 A ATOM 1412 C VAL A 189 18.240 46.610 −1.032 1.00 32.13A ATOM 1413 O VAL A 189 17.916 45.611 −1.660 1.00 31.84 A ATOM 1414 NLEU A 190 17.495 47.714 −1.005 1.00 32.08 A ATOM 1415 CA LEU A 19016.220 47.798 −1.718 1.00 32.32 A ATOM 1416 CB LEU A 190 15.539 49.139−1.426 1.00 33.11 A ATOM 1417 CG LEU A 190 14.876 49.287 −0.052 1.0032.87 A ATOM 1418 CD1 LEU A 190 14.378 50.724 0.120 1.00 33.09 A ATOM1419 CD2 LEU A 190 13.713 48.304 0.071 1.00 31.45 A ATOM 1420 C LEU A190 16.362 47.618 −3.229 1.00 33.42 A ATOM 1421 O LEU A 190 15.55546.936 −3.855 1.00 33.09 A ATOM 1422 N ALA A 191 17.379 48.238 −3.8161.00 33.70 A ATOM 1423 CA ALA A 191 17.600 48.113 −5.251 1.00 34.39 AATOM 1424 CB ALA A 191 18.766 48.987 −5.681 1.00 36.16 A ATOM 1425 C ALAA 191 17.886 46.656 −5.576 1.00 35.14 A ATOM 1426 O ALA A 191 17.39946.122 −6.572 1.00 36.27 A ATOM 1427 N LEU A 192 18.674 46.012 −4.7181.00 36.49 A ATOM 1428 CA LEU A 192 19.023 44.612 −4.902 1.00 34.74 AATOM 1429 CB LEU A 192 20.024 44.165 −3.830 1.00 34.60 A ATOM 1430 CGLEU A 192 20.375 42.671 −3.754 1.00 34.43 A ATOM 1431 CD1 LEU A 19220.924 42.176 −5.084 1.00 33.47 A ATOM 1432 CD2 LEU A 192 21.394 42.451−2.645 1.00 34.05 A ATOM 1433 C LEU A 192 17.782 43.727 −4.858 1.0035.29 A ATOM 1434 O LEU A 192 17.570 42.927 −5.767 1.00 33.59 A ATOM1435 N VAL A 193 16.962 43.865 −3.816 1.00 35.74 A ATOM 1436 CA VAL A193 15.763 43.039 −3.717 1.00 38.00 A ATOM 1437 CB VAL A 193 14.98543.274 −2.395 1.00 37.63 A ATOM 1438 CG1 VAL A 193 15.920 43.105 −1.2011.00 39.87 A ATOM 1439 CG2 VAL A 193 14.351 44.634 −2.389 1.00 37.77 AATOM 1440 C VAL A 193 14.837 43.327 −4.887 1.00 38.50 A ATOM 1441 O VALA 193 14.280 42.413 −5.483 1.00 38.16 A ATOM 1442 N ARG A 194 14.67744.604 −5.207 1.00 40.37 A ATOM 1443 CA ARG A 194 13.830 45.019 −6.3141.00 43.28 A ATOM 1444 CB ARG A 194 13.899 46.549 −6.461 1.00 45.33 AATOM 1445 CG ARG A 194 12.637 47.237 −6.974 1.00 50.72 A ATOM 1446 CDARG A 194 12.881 48.130 −8.183 1.00 54.41 A ATOM 1447 NE ARG A 19411.661 48.393 −8.942 1.00 57.37 A ATOM 1448 CZ ARG A 194 10.458 48.505−8.389 1.00 59.05 A ATOM 1449 NH1 ARG A 194 10.317 48.368 −7.079 1.0061.10 A ATOM 1450 NH2 ARG A 194 9.398 48.773 −9.140 1.00 61.23 A ATOM1451 C ARG A 194 14.283 44.305 −7.588 1.00 41.65 A ATOM 1452 O ARG A 19413.481 43.683 −8.280 1.00 41.71 A ATOM 1453 N GLU A 195 15.576 44.360−7.877 1.00 41.34 A ATOM 1454 CA GLU A 195 16.094 43.713 −9.070 1.0041.44 A ATOM 1455 CB GLU A 195 17.565 44.071 −9.264 1.00 43.69 A ATOM1456 CG GLU A 195 17.882 44.479 −10.693 1.00 46.31 A ATOM 1457 CD GLU A195 19.346 44.778 −10.911 1.00 45.36 A ATOM 1458 OE1 GLU A 195 19.87945.680 −10.233 1.00 45.90 A ATOM 1459 OE2 GLU A 195 19.960 44.107−11.765 1.00 46.65 A ATOM 1460 C GLU A 195 15.925 42.189 −9.052 1.0041.97 A ATOM 1461 O GLU A 195 15.700 41.577 −10.097 1.00 41.48 A ATOM1462 N LEU A 196 16.032 41.576 −7.874 1.00 40.20 A ATOM 1463 CA LEU A196 15.878 40.126 −7.762 1.00 38.53 A ATOM 1464 CB LEU A 196 16.68639.590 −6.570 1.00 36.27 A ATOM 1465 CG LEU A 196 18.212 39.731 −6.6491.00 37.41 A ATOM 1466 CD1 LEU A 196 18.852 39.207 −5.371 1.00 36.85 AATOM 1467 CD2 LEU A 196 18.740 38.969 −7.855 1.00 37.56 A ATOM 1468 CLEU A 196 14.399 39.753 −7.608 1.00 38.13 A ATOM 1469 O LEU A 196 14.04738.576 −7.509 1.00 38.58 A ATOM 1470 N GLY A 197 13.541 40.768 −7.5901.00 38.66 A ATOM 1471 CA GLY A 197 12.112 40.537 −7.458 1.00 38.24 AATOM 1472 C GLY A 197 11.712 39.891 −6.149 1.00 38.25 A ATOM 1473 O GLYA 197 10.833 39.028 −6.121 1.00 38.47 A ATOM 1474 N LEU A 198 12.33440.318 −5.055 1.00 37.09 A ATOM 1475 CA LEU A 198 12.031 39.733 −3.7561.00 36.88 A ATOM 1476 CB LEU A 198 13.340 39.324 −3.065 1.00 35.42 AATOM 1477 CG LEU A 198 14.267 38.410 −3.874 1.00 36.31 A ATOM 1478 CD1LEU A 198 15.612 38.284 −3.170 1.00 37.54 A ATOM 1479 CD2 LEU A 19813.631 37.037 −4.050 1.00 36.41 A ATOM 1480 C LEU A 198 11.211 40.629−2.827 1.00 36.62 A ATOM 1481 O LEU A 198 11.042 41.829 −3.064 1.0033.54 A ATOM 1482 N HIS A 199 10.707 40.025 −1.759 1.00 36.58 A ATOM1483 CA HIS A 199 9.918 40.738 −0.763 1.00 37.83 A ATOM 1484 CB HIS A199 9.667 39.833 0.440 1.00 39.68 A ATOM 1485 CG HIS A 199 8.795 40.4471.487 1.00 39.54 A ATOM 1486 CD2 HIS A 199 9.094 40.986 2.692 1.00 41.01A ATOM 1487 ND1 HIS A 199 7.429 40.549 1.348 1.00 42.14 A ATOM 1488 CE1HIS A 199 6.921 41.121 2.426 1.00 42.34 A ATOM 1489 NE2 HIS A 199 7.91141.396 3.257 1.00 43.77 A ATOM 1490 C HIS A 199 10.650 41.992 −0.2961.00 38.39 A ATOM 1491 O HIS A 199 11.845 41.950 0.012 1.00 38.41 A ATOM1492 N VAL A 200 9.928 43.107 −0.246 1.00 37.60 A ATOM 1493 CA VAL A 20010.506 44.367 0.188 1.00 37.98 A ATOM 1494 CB VAL A 200 9.689 45.569−0.348 1.00 38.84 A ATOM 1495 CG1 VAL A 200 10.254 46.869 0.201 1.0039.30 A ATOM 1496 CG2 VAL A 200 9.709 45.578 −1.878 1.00 39.44 A ATOM1497 C VAL A 200 10.537 44.447 1.714 1.00 38.23 A ATOM 1498 O VAL A 2009.506 44.353 2.369 1.00 37.38 A ATOM 1499 N PRO A 201 11.734 44.6042.302 1.00 38.40 A ATOM 1500 CD PRO A 201 13.070 44.490 1.687 1.00 36.38A ATOM 1501 CA PRO A 201 11.835 44.694 3.761 1.00 38.03 A ATOM 1502 CBPRO A 201 13.306 44.377 4.012 1.00 39.05 A ATOM 1503 CG PRO A 201 13.97144.949 2.792 1.00 37.38 A ATOM 1504 C PRO A 201 11.436 46.091 4.252 1.0039.00 A ATOM 1505 O PRO A 201 11.552 47.068 3.516 1.00 38.29 A ATOM 1506N ASN A 202 10.947 46.191 5.483 1.00 38.73 A ATOM 1507 CA ASN A 20210.564 47.492 6.007 1.00 38.53 A ATOM 1508 CB ASN A 202 9.273 47.3936.827 1.00 37.55 A ATOM 1509 CG ASN A 202 9.323 46.302 7.861 1.00 36.97A ATOM 1510 OD1 ASN A 202 10.368 46.051 8.463 1.00 34.21 A ATOM 1511 ND2ASN A 202 8.183 45.650 8.090 1.00 35.55 A ATOM 1512 C ASN A 202 11.68748.072 6.857 1.00 39.15 A ATOM 1513 O ASN A 202 12.798 47.539 6.877 1.0038.30 A ATOM 1514 N GLU A 203 11.392 49.161 7.561 1.00 38.76 A ATOM 1515CA GLU A 203 12.386 49.835 8.391 1.00 41.10 A ATOM 1516 CB GLU A 20311.727 50.944 9.212 1.00 44.04 A ATOM 1517 CG GLU A 203 12.689 52.0409.638 1.00 49.57 A ATOM 1518 CD GLU A 203 12.048 53.040 10.584 1.0052.76 A ATOM 1519 OE1 GLU A 203 10.873 53.402 10.362 1.00 54.28 A ATOM1520 OE2 GLU A 203 12.722 53.470 11.543 1.00 54.88 A ATOM 1521 C GLU A203 13.128 48.889 9.328 1.00 39.75 A ATOM 1522 O GLU A 203 14.353 48.9419.434 1.00 38.21 A ATOM 1523 N LEU A 204 12.383 48.028 10.006 1.00 38.08A ATOM 1524 CA LEU A 204 12.984 47.081 10.932 1.00 37.43 A ATOM 1525 CBLEU A 204 11.884 46.297 11.649 1.00 39.35 A ATOM 1526 CG LEU A 20412.342 45.230 12.644 1.00 41.12 A ATOM 1527 CD1 LEU A 204 13.153 45.88213.753 1.00 41.94 A ATOM 1528 CD2 LEU A 204 11.131 44.512 13.218 1.0043.55 A ATOM 1529 C LEU A 204 13.922 46.121 10.195 1.00 37.20 A ATOM1530 O LEU A 204 15.060 45.897 10.624 1.00 36.41 A ATOM 1531 N GLY A 20513.438 45.561 9.086 1.00 34.65 A ATOM 1532 CA GLY A 205 14.240 44.6348.303 1.00 35.07 A ATOM 1533 C GLY A 205 15.512 45.257 7.756 1.00 34.00A ATOM 1534 O GLY A 205 16.577 44.628 7.753 1.00 33.65 A ATOM 1535 N LEUA 206 15.408 46.500 7.297 1.00 34.66 A ATOM 1536 CA LEU A 206 16.55947.201 6.734 1.00 35.09 A ATOM 1537 CB LEU A 206 16.095 48.464 6.01213.00 36.03 A ATOM 1538 CG LEU A 206 15.387 48.179 4.679 1.00 33.49 AATOM 1539 CD1 LEU A 206 14.670 49.426 4.182 1.00 33.05 A ATOM 1540 CD2LEU A 206 16.404 47.701 3.662 1.00 32.40 A ATOM 1541 C LEU A 206 17.61647.532 7.782 1.00 36.87 A ATOM 1542 O LEU A 206 18.821 47.468 7.505 1.0036.00 A ATOM 1543 N LYS A 207 17.168 47.878 8.987 1.00 36.83 A ATOM 1544CA LYS A 207 18.091 48.177 10.074 1.00 37.90 A ATOM 1545 CB LYS A 20717.320 48.600 11.326 1.00 39.99 A ATOM 1546 CG LYS A 207 18.216 49.00712.495 1.00 45.04 A ATOM 1547 CD LYS A 207 17.400 49.347 13.740 1.0050.01 A ATOM 1548 CE LYS A 207 18.268 49.988 14.828 1.00 52.95 A ATOM1549 NZ LYS A 207 19.403 49.110 15.254 1.00 55.35 A ATOM 1550 C LYS A207 18.891 46.903 10.362 1.00 37.50 A ATOM 1551 O LYS A 207 20.09546.951 10.611 1.00 38.54 A ATOM 1552 N PHE A 208 18.208 45.762 10.3121.00 35.87 A ATOM 1553 CA PHE A 208 18.831 44.468 10.552 1.00 35.98 AATOM 1554 CB PHE A 208 17.746 43.376 10.613 1.00 34.44 A ATOM 1555 CGPHE A 208 18.288 41.966 10.743 1.00 36.93 A ATOM 1556 CD1 PHE A 20818.732 41.264 9.621 1.00 35.75 A ATOM 1557 CD2 PHE A 208 18.328 41.33211.987 1.00 36.14 A ATOM 1558 CE1 PHE A 208 19.201 39.955 9.734 1.0035.55 A ATOM 1559 CE2 PHE A 208 18.798 40.020 12.110 1.00 37.86 A ATOM1560 CZ PHE A 208 19.234 39.330 10.977 1.00 37.36 A ATOM 1561 C PHE A208 19.866 44.149 9.464 1.00 35.26 A ATOM 1562 O PHE A 208 20.576 43.7009.758 1.00 34.23 A ATOM 1563 N CYS A 209 19.508 44.389 8.208 1.00 35.13A ATOM 1564 CA CYS A 209 20.424 44.115 7.106 1.00 35.37 A ATOM 1565 CBCYS A 209 19.756 44.438 5.762 1.00 33.36 A ATOM 1566 SG CYS A 209 18.35743.364 5.322 1.00 33.51 A ATOM 1567 C CYS A 209 21.720 44.920 7.244 1.0035.58 A ATOM 1568 O CYS A 209 22.805 44.416 6.961 1.00 36.94 A ATOM 1569N LYS A 210 21.592 46.172 7.669 1.00 35.97 A ATOM 1570 CA LYS A 21022.736 47.067 7.844 1.00 36.87 A ATOM 1571 CB LYS A 210 22.239 48.4298.341 1.00 37.10 A ATOM 1572 CG LYS A 210 23.296 49.330 8.973 1.00 42.31A ATOM 1573 CD LYS A 210 22.677 50.663 9.409 1.00 44.14 A ATOM 1574 CELYS A 210 23.705 51.801 9.307 1.00 47.02 A ATOM 1575 NZ LYS A 210 23.11453.178 9.361 1.00 48.89 A ATOM 1576 C LYS A 210 23.761 46.497 8.824 1.0036.21 A ATOM 1577 O LYS A 210 24.949 46.793 8.739 1.00 37.65 A ATOM 1578N ARG A 211 23.285 45.669 9.743 1.00 35.86 A ATOM 1579 CA ARG A 21124.123 45.053 10.765 1.00 36.74 A ATOM 1580 CB ARG A 211 23.273 44.86312.023 1.00 37.38 A ATOM 1581 CG ARG A 211 23.965 44.280 13.223 1.0040.40 A ATOM 1582 CD ARG A 211 23.039 44.393 14.423 1.00 43.85 A ATOM1583 NE ARG A 211 23.632 43.875 15.651 1.00 46.63 A ATOM 1584 CZ ARG A211 23.104 44.050 16.857 1.00 48.65 A ATOM 1585 NH1 ARG A 211 21.97344.734 16.992 1.00 49.90 A ATOM 1586 NH2 ARG A 211 23.697 43.535 17.9291.00 48.82 A ATOM 1587 C ARG A 211 24.735 43.715 10.325 1.00 35.55 AATOM 1588 O ARG A 211 25.559 43.141 11.038 1.00 35.83 A ATOM 1589 N SERA 212 24.345 45.238 9.144 1.00 33.22 A ATOM 1590 CA SER A 212 24.81841.954 8.607 1.00 33.87 A ATOM 1591 CB SER A 212 24.125 41.652 7.2691.00 31.55 A ATOM 1592 OG SER A 212 22.738 41.403 7.448 1.00 33.21 AATOM 1593 C SER A 212 26.319 41.789 8.401 1.00 32.15 A ATOM 1594 O SER A212 27.009 42.714 8.009 1.00 35.39 A ATOM 1595 N PHE A 213 26.818 40.5858.657 1.00 33.61 A ATOM 1596 CA PHE A 213 28.236 40.295 8.447 1.00 34.25A ATOM 1597 CB PHE A 213 28.862 39.641 9.695 1.00 33.84 A ATOM 1598 CGPHE A 213 28.500 38.192 9.887 1.00 34.55 A ATOM 1599 CD1 PHE A 21329.384 37.186 9.509 1.00 34.18 A ATOM 1600 CD2 PHE A 213 27.291 37.83410.471 1.00 34.89 A ATOM 1601 CE1 PHE A 213 29.071 35.846 9.717 1.0037.02 A ATOM 1602 CE2 PHE A 213 26.965 36.501 10.683 1.00 36.59 A ATOM1603 CZ PHE A 213 27.858 35.500 10.306 1.00 35.85 A ATOM 1604 C PHE A213 28.349 39.363 7.242 1.00 33.28 A ATOM 1605 O PHE A 213 29.422 39.1786.685 1.00 32.96 A ATOM 1606 N SER A 214 27.222 38.786 6.838 1.00 34.50A ATOM 1607 CA SER A 214 27.192 37.874 5.698 1.00 31.86 A ATOM 1608 CBSER A 214 27.375 36.434 6.170 1.00 33.79 A ATOM 1609 OG SER A 214 27.22435.521 5.093 1.00 37.93 A ATOM 1610 C SER A 214 25.860 37.988 4.974 1.0031.77 A ATOM 1611 O SER A 214 24.833 38.202 5.610 1.00 28.63 A ATOM 1612N VAL A 215 25.892 37.856 3.648 1.00 30.90 A ATOM 1613 CA VAL A 21524.685 37.908 2.818 1.00 29.86 A ATOM 1614 CB VAL A 215 24.618 39.1871.947 1.00 30.29 A ATOM 1615 CG1 VAL A 215 23.443 39.103 0.993 1.0032.52 A ATOM 1616 CG2 VAL A 215 24.460 40.412 2.817 1.00 33.53 A ATOM1617 C VAL A 215 24.759 36.725 1.866 1.00 28.93 A ATOM 1618 O VAL A 21525.802 36.513 1.238 1.00 28.44 A ATOM 1619 N TYR A 216 23.679 35.9521.749 1.00 27.25 A ATOM 1620 CA TYR A 216 23.703 34.817 0.824 1.00 27.51A ATOM 1621 CB TYR A 216 24.119 33.499 1.522 1.00 26.91 A ATOM 1622 CGTYR A 216 23.361 33.072 2.774 1.00 27.45 A ATOM 1623 CD1 TYR A 21622.508 31.959 2.769 1.00 26.55 A ATOM 1624 CE1 TYR A 216 21.913 31.4873.967 1.00 29.46 A ATOM 1625 CD2 TYR A 216 23.593 33.710 3.992 1.0030.60 A ATOM 1626 CE2 TYR A 216 23.016 33.258 5.178 1.00 29.33 A ATOM1627 CZ TYR A 216 22.186 32.156 5.165 1.00 29.32 A ATOM 1628 OH TYR A216 21.654 31.734 6.360 1.00 31.96 A ATOM 1629 C TYR A 216 22.477 34.534−0.019 1.00 27.43 A ATOM 1630 O TYR A 216 21.383 34.337 0.492 1.00 25.60A ATOM 1631 N PRO A 217 22.658 34.519 −1.349 1.00 28.06 A ATOM 1632 CDPRO A 217 23.839 35.061 −2.054 1.00 28.05 A ATOM 1633 CA PRO A 21721.576 34.241 −2.293 1.00 27.27 A ATOM 1634 CB PRO A 217 22.008 35.008−3.534 1.00 28.73 A ATOM 1635 CG PRO A 217 23.512 34.790 −3.521 1.0027.12 A ATOM 1636 C PRO A 217 21.558 32.736 −2.550 1.00 27.30 A ATOM1637 O PRO A 217 22.596 32.066 −2.432 1.00 26.63 A ATOM 1638 N THR A 21820.393 32.204 −2.890 1.00 25.62 A ATOM 1639 CA THR A 218 20.258 30.784−3.204 1.00 26.97 A ATOM 1640 CB THR A 218 19.073 30.132 −2.452 1.0027.35 A ATOM 1641 OG1 THR A 218 19.274 30.242 −1.035 1.00 28.53 A ATOM1642 CG2 THR A 218 18.962 28.645 −2.818 1.00 26.30 A ATOM 1643 C THR A218 19.991 30.674 −4.707 1.00 28.41 A ATOM 1644 O THR A 218 19.11531.364 −5.231 1.00 29.31 A ATOM 1645 N LEU A 219 20.755 29.823 −5.3881.00 29.45 A ATOM 1646 CA LEU A 219 20.608 29.619 −6.826 1.00 31.35 AATOM 1647 CB LEU A 219 21.884 30.066 −7.562 1.00 32.22 A ATOM 1648 CGLEU A 219 22.337 31.509 −7.312 1.00 32.16 A ATOM 1649 CD1 LEU A 21923.346 31.516 −6.160 1.00 31.96 A ATOM 1650 CD2 LEU A 219 22.961 32.105−8.584 1.00 29.71 A ATOM 1651 C LEU A 219 20.315 28.149 −7.148 1.0032.09 A ATOM 1652 O LEU A 219 20.540 27.255 −6.239 1.00 31.23 A ATOM1653 N ASN A 220 19.812 27.898 −8.349 1.00 33.47 A ATOM 1654 CA ASN A220 19.498 26.537 −8.767 1.00 34.71 A ATOM 1655 CB ASN A 220 17.99026.289 −8.648 1.00 36.03 A ATOM 1656 CG ASN A 220 17.186 27.115 −9.6291.00 39.61 A ATOM 1657 OD1 ASN A 220 17.141 26.809 −10.819 1.00 40.74 AATOM 1658 ND2 ASN A 220 16.558 28.177 −9.138 1.00 38.72 A ATOM 1659 CASN A 220 19.965 26.294 −10.205 1.00 35.49 A ATOM 1660 O ASN A 22020.354 27.228 −10.906 1.00 33.83 A ATOM 1661 N TRP A 221 19.927 25.033−10.623 1.00 36.36 A ATOM 1662 CA TRP A 221 20.354 24.615 −11.956 1.0038.77 A ATOM 1663 CB TRP A 221 20.845 23.162 −11.906 1.00 37.13 A ATOM1664 CG TRP A 221 22.255 22.982 −11.415 1.00 35.49 A ATOM 1665 CD2 TRP A221 22.679 22.785 −10.056 1.00 34.23 A ATOM 1666 CE2 TRP A 221 24.08422.613 −10.078 1.00 34.61 A ATOM 1667 CE3 TRP A 221 22.009 22.733 −8.8261.00 33.47 A ATOM 1668 CD1 TRP A 221 23.388 22.932 −12.180 1.00 36.61 AATOM 1669 NE1 TRP A 221 24.490 22.707 −11.384 1.00 36.62 A ATOM 1670 CZ2TRP A 221 24.832 22.391 −8.916 1.00 31.63 A ATOM 1671 CZ3 TRP A 22122.752 22.511 −7.665 1.00 31.82 A ATOM 1672 CH2 TRP A 221 24.153 22.343−7.722 1.00 33.18 A ATOM 1673 C TRP A 221 19.261 24.736 −13.020 1.0041.28 A ATOM 1674 O TRP A 221 19.546 24.642 −14.214 1.00 41.24 A ATOM1675 N GLU A 222 18.017 24.943 −12.954 1.00 42.94 A ATOM 1676 CA GLU A222 16.905 25.058 −13.534 1.00 46.16 A ATOM 1677 CB GLU A 222 15.58724.689 −12.848 1.00 46.48 A ATOM 1678 CG GLU A 222 15.530 23.273 −12.3031.00 49.88 A ATOM 1679 CD GLU A 222 15.994 22.238 −13.307 1.00 52.45 AATOM 1680 OE1 GLU A 222 15.705 22.404 −14.512 1.00 55.71 A ATOM 1681 OE2GLU A 222 16.640 21.251 −12.894 1.00 53.80 A ATOM 1682 C GLU A 22216.748 26.425 −14.203 1.00 46.68 A ATOM 1683 O GLU A 222 16.463 26.501−15.398 1.00 47.48 A ATOM 1684 N THR A 223 16.920 27.500 −13.439 1.0046.30 A ATOM 1685 CA THR A 223 16.777 28.848 −13.985 1.00 46.13 A ATOM1686 CB THR A 223 15.407 29.462 −13.613 1.00 47.72 A ATOM 1687 OG1 THR A223 15.332 29.646 −12.194 1.00 49.52 A ATOM 1688 CG2 THR A 223 14.27028.546 −14.051 1.00 49.10 A ATOM 1689 C THR A 223 17.867 29.791 −13.4771.00 46.02 A ATOM 1690 O THR A 223 18.643 29.437 −12.591 1.00 44.84 AATOM 1691 N GLY A 224 17.915 30.994 −14.044 1.00 44.46 A ATOM 1692 CAGLY A 224 18.906 31.971 −13.626 1.00 43.49 A ATOM 1693 C GLY A 22418.341 32.872 −12.545 1.00 42.65 A ATOM 1694 O GLY A 224 19.014 33.777−12.057 1.00 43.27 A ATOM 1695 N LYS A 225 17.093 32.618 −12.167 1.0041.43 A ATOM 1696 CA LYS A 225 16.426 33.415 −11.150 1.00 41.27 A ATOM1697 CB LYS A 225 14.908 33.310 −11.320 1.00 43.63 A ATOM 1698 CG LYS A225 14.111 34.119 −10.308 1.00 47.97 A ATOM 1699 CD LYS A 225 12.60433.860 −10.427 1.00 52.08 A ATOM 1700 CE LYS A 225 12.243 32.430 −10.0171.00 53.90 A ATOM 1701 NZ LYS A 225 10.779 32.155 −10.145 1.00 56.20 AATOM 1702 C LYS A 225 16.816 32.981 −9.734 1.00 38.89 A ATOM 1703 O LYSA 225 16.693 31.812 −9.376 1.00 36.62 A ATOM 1704 N ILE A 226 17.29033.934 −8.939 1.00 37.52 A ATOM 1705 CA ILE A 226 17.676 33.648 −7.5651.00 35.91 A ATOM 1706 CB ILE A 226 18.415 34.860 −6.939 1.00 34.66 AATOM 1707 CG2 ILE A 226 18.474 34.725 −5.412 1.00 32.77 A ATOM 1708 CG1ILE A 226 19.831 34.932 −7.526 1.00 35.67 A ATOM 1709 CD1 ILE A 22620.616 36.173 −7.142 1.00 35.94 A ATOM 1710 C ILE A 226 16.419 33.297−6.775 1.00 35.16 A ATOM 1711 O ILE A 226 15.420 34.010 −6.838 1.0036.27 A ATOM 1712 N ASP A 227 16.472 32.183 −6.048 1.00 34.90 A ATOM1713 CA ASP A 227 15.337 31.700 −5.260 1.00 35.04 A ATOM 1714 CB ASP A227 15.589 30.250 −4.845 1.00 37.48 A ATOM 1715 CG ASP A 227 15.76829.329 −6.041 1.00 40.26 A ATOM 1716 OD1 ASP A 227 14.795 29.161 −6.8111.00 40.93 A ATOM 1717 OD2 ASP A 227 16.881 28.784 −6.213 1.00 40.47 AATOM 1718 C ASP A 227 15.009 32.536 −4.024 1.00 35.52 A ATOM 1719 O ASPA 227 13.836 32.772 −3.710 1.00 33.67 A ATOM 1720 N ARG A 228 16.04532.966 −3.311 1.00 33.10 A ATOM 1721 CA ARG A 228 15.859 33.774 −2.1171.00 31.73 A ATOM 1722 CB ARG A 228 15.297 32.910 −0.974 1.00 31.32 AATOM 1723 CG ARG A 228 16.153 31.704 −0.621 1.00 29.51 A ATOM 1724 CDARG A 228 15.468 30.793 0.405 1.00 32.46 A ATOM 1725 NE ARG A 228 16.37029.723 0.826 1.00 31.71 A ATOM 1726 CZ ARG A 228 16.486 28.550 0.2151.00 33.70 A ATOM 1727 NH1 ARG A 228 15.741 28.266 −0.848 1.00 35.15 AATOM 1728 NH2 ARG A 228 17.382 27.675 0.646 1.00 35.72 A ATOM 1729 C ARGA 228 17.189 34.408 −1.723 1.00 30.83 A ATOM 1730 O ARG A 228 18.23934.052 −2.264 1.00 32.29 A ATOM 1731 N LEU A 229 17.134 34.353 −0.7931.00 31.06 A ATOM 1732 CA LEU A 229 18.327 36.066 −0.330 1.00 29.02 AATOM 1733 CB LEU A 229 18.365 37.451 −0.964 1.00 27.74 A ATOM 1734 CGLEU A 229 19.548 38.344 −0.595 1.00 26.58 A ATOM 1735 CD1 LEU A 22920.822 37.775 −1.210 1.00 24.82 A ATOM 1736 CD2 LEU A 229 19.266 39.765−1.083 1.00 26.37 A ATOM 1737 C LEU A 229 18.293 36.211 1.187 1.00 27.73A ATOM 1738 O LEU A 229 17.310 36.701 1.743 1.00 28.78 A ATOM 1739 N CYSA 230 19.367 35.808 1.855 1.00 28.33 A ATOM 1740 CA CYS A 230 19.40435.894 3.313 1.00 29.29 A ATOM 1741 CB LYS A 230 19.510 34.485 3.9091.00 29.74 A ATOM 1742 SG CYS A 230 19.579 34.453 5.743 1.00 31.41 AATOM 1743 C CYS A 230 20.523 36.777 3.878 1.00 28.54 A ATOM 1744 O CYS A230 21.661 36.749 3.406 1.00 28.22 A ATOM 1745 N PHE A 231 20.172 37.5744.882 1.00 28.82 A ATOM 1746 CA PHE A 231 21.114 38.451 5.564 1.00 29.99A ATOM 1747 CB PHE A 231 20.539 39.687 5.695 1.00 28.76 A ATOM 1748 CGPHE A 231 20.349 40.570 4.371 1.00 31.79 A ATOM 1749 CD1 PHE A 23119.261 40.265 3.550 1.00 31.62 A ATOM 1750 CD2 PHE A 231 21.279 41.5153.930 1.00 31.11 A ATOM 1751 CE1 PHE A 231 19.103 40.891 2.305 1.0030.59 A ATOM 1752 CE2 PHE A 231 21.130 42.147 2.683 1.00 31.97 A ATOM1753 CZ PHE A 231 20.038 41.830 1.873 1.00 29.20 A ATOM 1754 C PHE A 23121.339 37.846 6.946 1.00 30.94 A ATOM 1755 O PHE A 231 20.380 37.5537.660 1.00 30.10 A ATOM 1756 N ALA A 232 22.600 37.645 7.319 1.00 30.85A ATOM 1757 CA ALA A 232 22.916 37.051 8.617 1.00 31.42 A ATOM 1758 CBALA A 232 23.814 35.825 8.420 1.00 30.46 A ATOM 1759 C ALA A 232 23.58438.039 9.580 1.00 30.98 A ATOM 1760 O ALA A 232 24.402 38.857 9.175 1.0031.61 A ATOM 1761 N VAL A 233 23.227 37.933 10.856 1.00 32.02 A ATOM1762 CA VAL A 233 23.757 38.794 11.911 1.00 33.05 A ATOM 1763 CB VAL A233 22.651 39.748 12.449 1.00 34.08 A ATOM 1764 CG1 VAL A 233 23.10940.432 13.753 1.00 35.86 A ATOM 1765 CG2 VAL A 233 22.325 40.800 11.3961.00 31.79 A ATOM 1766 C VAL A 233 24.271 37.931 13.065 1.00 33.07 AATOM 1767 O VAL A 233 23.580 37.024 13.524 1.00 34.33 A ATOM 1768 N ILEA 234 25.485 38.211 13.526 1.00 34.06 A ATOM 1769 CA ILE A 234 26.07337.449 14.623 1.00 36.42 A ATOM 1770 CB ILE A 234 27.519 37.004 14.2601.00 34.49 A ATOM 1771 CG2 ILE A 234 28.420 38.209 14.104 1.00 35.27 AATOM 1772 CG1 ILE A 234 28.051 36.032 15.316 1.00 35.08 A ATOM 1773 CD1ILE A 234 29.314 35.277 14.888 1.00 32.27 A ATOM 1774 C ILE A 234 26.05338.313 15.888 1.00 35.82 A ATOM 1775 O ILE A 234 26.473 39.461 15.8631.00 36.21 A ATOM 1776 N SER A 235 25.549 37.772 16.989 1.00 38.61 AATOM 1777 CA SER A 235 25.471 38.561 18.219 1.00 40.77 A ATOM 1778 CBSER A 235 24.385 39.633 18.065 1.00 41.97 A ATOM 1779 OG SER A 23524.220 40.392 19.248 1.00 43.56 A ATOM 1780 C SER A 235 25.175 37.74219.467 1.00 42.35 A ATOM 1781 O SER A 235 24.793 36.574 19.384 1.0041.79 A ATOM 1782 N ASN A 236 25.356 38.372 20.626 1.00 44.38 a ATOM1783 CA ASN A 236 25.081 37.731 21.906 1.00 45.74 A ATOM 1784 CB ASN A236 26.115 38.151 22.958 1.00 47.60 A ATOM 1785 CG ASN A 236 27.48837.569 22.694 1.00 48.75 A ATOM 1786 OD1 ASN A 236 27.661 36.350 22.6801.00 51.03 A ATOM 1787 ND2 ASN A 236 28.472 38.436 22.479 1.00 49.64 AATOM 1788 C ASN A 236 23.692 38.172 22.350 1.00 46.12 A ATOM 1789 O ASNA 236 23.088 37.568 23.236 1.00 47.30 A ATOM 1790 N ASP A 237 23.20139.230 21.713 1.00 46.49 A ATOM 1791 CA ASP A 237 21.889 39.806 21.9931.00 46.83 A ATOM 1792 CB ASP A 237 21.746 41.105 21.198 1.00 47.75 AATOM 1793 CG ASP A 237 20.662 42.013 21.736 1.00 47.79 A ATOM 1794 OD1ASP A 237 19.513 41.560 21.923 1.00 47.65 A ATOM 1795 OD2 ASP A 23720.970 43.198 21.957 1.00 48.75 A ATOM 1796 C ASP A 237 20.786 38.82521.591 1.00 47.10 A ATOM 1797 O ASP A 237 20.665 38.463 20.422 1.0047.73 A ATOM 1798 N PRO A 238 19.965 38.383 22.557 1.00 46.57 A ATOM1799 CD PRO A 238 20.211 38.488 24.009 1.00 47.58 A ATOM 1800 CA PRO A238 18.878 37.439 22.281 1.00 45.52 A ATOM 1801 CB PRO A 238 18.79636.641 23.576 1.00 47.17 A ATOM 1802 CG PRO A 238 19.055 37.703 24.6051.00 47.42 A ATOM 1803 C PRO A 238 17.530 38.049 21.893 1.00 45.24 AATOM 1804 O PRO A 238 16.509 37.361 21.900 1.00 45.57 A ATOM 1805 N THRA 239 17.521 39.331 21.545 1.00 45.54 A ATOM 1806 CA THR A 239 16.28040.001 21.153 1.00 45.56 A ATOM 1807 CB THR A 239 15.920 41.117 22.1621.00 45.90 A ATOM 1808 OG1 THR A 239 16.886 42.173 22.067 1.00 46.13 AATOM 1809 CG2 THR A 239 15.921 40.574 23.588 1.00 46.71 A ATOM 1810 CTHR A 239 16.372 40.642 19.761 1.00 44.94 A ATOM 1811 O THR A 239 15.65741.600 19.473 1.00 44.48 A ATOM 1812 N LEU A 240 17.226 40.106 18.8921.00 45.21 A ATOM 1813 CA LEU A 240 17.412 40.689 17.563 1.00 44.68 AATOM 1814 CB LEU A 240 18.887 40.606 17.167 1.00 44.71 A ATOM 1815 CGLEU A 240 19.890 41.396 18.008 1.00 45.86 A ATOM 1816 CD1 LEU A 24021.292 41.136 17.488 1.00 46.25 A ATOM 1817 CD2 LEU A 240 19.566 42.88517.952 1.00 45.57 A ATOM 1818 C LEU A 240 16.573 40.190 16.387 1.0044.93 A ATOM 1819 O LEU A 240 16.801 40.625 15.258 1.00 44.99 A ATOM1820 N VAL A 241 15.614 32.298 16.617 1.00 44.69 A ATOM 1821 CA VAL A241 14.809 38.806 15.504 1.00 44.20 A ATOM 1822 CB VAL A 241 13.77537.744 15.976 1.00 44.62 A ATOM 1823 CG1 VAL A 241 12.661 38.393 16.7811.00 45.74 A ATOM 1824 CG2 VAL A 241 13.219 36.998 14.778 1.00 43.96 AATOM 1825 C VAL A 241 14.114 40.004 14.839 1.00 44.51 A ATOM 1826 O VALA 241 13.424 40.760 15.498 1.00 44.10 A ATOM 1827 N PRO A 242 14.30240.175 13.520 1.00 42.66 A ATOM 1828 CD PRO A 242 15.176 39.385 12.6351.00 41.53 A ATOM 1829 CA PRO A 242 13.696 41.293 12.787 1.00 41.75 AATOM 1830 CB PRO A 242 14.584 41.403 11.557 1.00 41.67 A ATOM 1831 CGPRO A 242 14.865 39.966 11.257 1.00 41.45 A ATOM 1832 C PRO A 242 12.22941.111 12.419 1.00 42.06 A ATOM 1833 O PRO A 242 11.849 41.287 11.2651.00 39.87 A ATOM 1834 N SER A 243 11.409 40.772 13.407 1.00 42.14 AATOM 1835 CA SER A 243 9.987 40.566 13.181 1.00 43.50 A ATOM 1836 CB SERA 243 9.629 39.096 13.421 1.00 42.47 A ATOM 1837 OG SER A 243 8.23738.875 13.288 1.00 41.75 A ATOM 1838 C SER A 243 9.170 41.460 14.1091.00 45.15 A ATOM 1839 O SER A 243 9.611 41.799 15.208 1.00 42.95 A ATOM1840 N SER A 244 7.981 41.844 13.657 1.00 47.40 A ATOM 1841 CA SER A 2447.102 42.697 14.450 1.00 49.90 A ATOM 1842 CB SER A 244 6.520 43.81313.579 1.00 49.67 A ATOM 1843 OG SER A 244 5.769 43.277 12.500 1.0048.66 A ATOM 1844 C SER A 244 5.971 41.865 15.033 1.00 51.74 A ATOM 1845O SER A 244 5.122 42.375 15.765 1.00 53.20 A ATOM 1846 N ASP A 245 5.96440.580 14.693 1.00 53.53 A ATOM 1847 CA ASP A 245 4.496 39.653 15.1711.00 55.50 A ATOM 1848 CB ASP A 245 4.823 38.470 14.207 1.00 56.42 AATOM 1849 CG ASP A 245 3.803 37.450 14.665 1.00 57.31 A ATOM 1850 OD1ASP A 245 2.631 37.830 14.862 1.00 59.97 A ATOM 1851 OD2 ASP A 245 4.17036.269 14.827 1.00 57.82 A ATOM 1852 C ASP A 245 5.283 39.149 16.5711.00 55.95 A ATOM 1853 O ASP A 245 6.250 38.411 16.758 1.00 55.39 A ATOM1854 N GLU A 246 4.479 39.562 17.549 1.00 57.53 A ATOM 1855 CA GLU A 2464.673 39.162 18.914 1.00 58.27 A ATOM 1856 CB GLU A 246 3.443 39.53719.776 1.00 60.44 A ATOM 1857 CG GLU A 246 3.265 41.033 19.999 1.0063.33 A ATOM 1858 CD GLU A 246 2.015 41.367 20.806 1.00 65.68 A ATOM1859 OE1 GLU A 246 0.893 41.126 20.306 1.00 65.94 A ATOM 1860 OE2 GLU A246 2.156 41.868 21.944 1.00 67.06 A ATOM 1861 C GLU A 246 4.949 37.66619.072 1.00 57.75 A ATOM 1862 O GLU A 246 5.645 37.236 19.989 1.00 57.64A ATOM 1863 N GLY A 247 4.400 36.876 18.155 1.00 57.63 A ATOM 1864 CAGLY A 247 4.625 35.440 18.196 1.00 57.31 A ATOM 1865 C GLY A 247 6.09735.098 18.032 1.00 56.48 A ATOM 1866 O GLY A 247 6.657 34.325 18.8141.00 56.42 A ATOM 1867 N ASP A 248 6.731 35.677 17.016 1.00 55.47 A ATOM1868 CA ASP A 248 8.146 35.430 16.760 1.00 54.34 A ATOM 1869 CB ASP A248 8.576 36.044 15.426 1.00 52.59 A ATOM 1870 CG ASP A 248 7.876 35.42714.246 1.00 54.48 A ATOM 1871 OD1 ASP A 248 7.731 34.188 14.225 1.0051.37 A ATOM 1872 OD2 ASP A 248 7.485 36.180 13.328 1.00 51.45 A ATOM1873 C ASP A 248 9.018 36.022 17.855 1.00 54.31 A ATOM 1874 O ASP A 2489.865 35.339 18.428 1.00 54.40 A ATOM 1875 N ILE A 249 8.813 37.30618.123 1.00 55.01 A ATOM 1876 CA ILE A 249 9.578 38.028 19.131 1.0055.17 A ATOM 1877 CB ILE A 249 8.906 39.378 19.454 1.00 55.80 A ATOM1878 CG2 ILE A 249 9.831 40.234 20.307 1.00 55.95 A ATOM 1879 CG1 ILE A249 8.575 40.109 18.151 1.00 56.11 A ATOM 1880 CD1 ILE A 249 7.74941.366 18.338 1.00 57.14 A ATOM 1881 C ILE A 249 9.682 37.201 20.4041.00 55.78 A ATOM 1882 O ILE A 249 10.697 37.234 21.102 1.00 55.95 AATOM 1883 N GLU A 250 8.626 36.450 20.690 1.00 55.60 A ATOM 1884 CA GLUA 250 8.577 35.611 21.875 1.00 55.90 A ATOM 1885 CB GLU A 250 7.12635.267 22.214 1.00 57.52 A ATOM 1886 CG GLU A 250 6.987 34.358 23.4161.00 60.65 A ATOM 1887 CD GLU A 250 7.686 34.919 24.634 1.00 62.45 AATOM 1888 OE1 GLU A 250 7.352 36.054 25.038 1.00 63.26 A ATOM 1889 OE2GLU A 250 8.572 34.229 25.182 1.00 63.88 A ATOM 1890 C GLU A 250 9.36834.323 21.698 1.00 54.72 A ATOM 1891 O GLU A 250 10.349 34.080 22.4051.00 53.29 A ATOM 1892 N LYS A 251 8.928 33.506 20.745 1.00 54.03 A ATOM1893 CA LYS A 251 9.555 32.221 20.461 1.00 53.48 A ATOM 1894 CB LYS A251 8.880 31.573 19.251 1.00 54.33 A ATOM 1895 CG LYS A 251 9.268 30.12219.033 1.00 55.38 A ATOM 1896 CD LYS A 251 8.414 29.489 17.953 1.0057.26 A ATOM 1897 CE LYS A 251 8.646 27.989 17.873 1.00 58.08 A ATOM1898 NZ LYS A 251 7.790 27.348 16.833 1.00 58.69 A ATOM 1899 C LYS A 25111.067 32.292 20.235 1.00 52.15 A ATOM 1900 O LYS A 251 11.812 31.44120.731 1.00 51.69 A ATOM 1901 N PHE A 252 11.523 33.300 19.497 1.0050.10 A ATOM 1902 CA PHE A 252 12.951 33.434 19.232 1.00 48.89 A ATOM1903 CB PHE A 252 13.197 34.435 18.099 1.00 45.87 A ATOM 1904 CG PHE A252 12.915 33.875 16.733 1.00 43.99 A ATOM 1905 CD1 PHE A 252 11.61633.564 16.349 1.00 41.90 A ATOM 1906 CD2 PHE A 252 13.957 33.623 15.8411.00 42.23 A ATOM 1907 CE1 PHE A 252 11.356 33.009 15.097 1.00 41.89 AATOM 1908 CE2 PHE A 252 13.708 33.069 14.587 1.00 41.06 A ATOM 1909 CZPHE A 252 12.409 32.762 14.214 1.00 41.75 A ATOM 1910 C PHE A 252 13.74433.837 20.466 1.00 48.68 A ATOM 1911 O PHE A 252 14.855 33.355 20.6801.00 49.06 A ATOM 1912 N HIS A 253 13.181 34.722 21.279 1.00 48.76 AATOM 1913 CA HIS A 253 13.870 35.145 22.490 1.00 49.30 A ATOM 1914 CBHIS A 253 13.079 36.237 23.215 1.00 49.55 A ATOM 1915 CG HIS A 25313.576 36.513 24.599 1.00 50.62 A ATOM 1916 CD2 HIS A 253 14.564 37.32425.045 1.00 50.93 A ATOM 1917 ND1 HIS A 253 13.084 35.861 25.709 1.0051.60 A ATOM 1918 CE1 HIS A 253 13.749 36.258 26.779 1.00 52.53 A ATOM1919 NE2 HIS A 253 14.653 37.144 26.404 1.00 52.16 A ATOM 1920 C HIS A253 14.033 33.937 23.403 1.00 48.72 A ATOM 1921 O HIS A 253 15.06733.765 24.052 1.00 47.39 A ATOM 1922 N ASN A 254 12.995 33.108 23.4371.00 48.76 A ATOM 1923 CA ASN A 254 12.986 31.900 24.246 1.00 50.80 AATOM 1924 CB ASN A 254 11.657 31.165 24.060 1.00 52.29 A ATOM 1925 CGASN A 254 11.531 29.948 24.955 1.00 55.40 A ATOM 1926 OD1 ASN A 25411.461 30.066 26.181 1.00 57.62 A ATOM 1927 ND2 ASN A 254 11.503 28.76824.345 1.00 55.81 A ATOM 1928 C ASN A 254 14.137 30.998 23.817 1.0050.81 A ATOM 1929 O ASN A 254 15.008 30.646 24.623 1.00 50.82 A ATOM1930 N TYR A 255 14.138 30.637 22.537 1.00 50.64 A ATOM 1931 CA TYR A255 15.171 29.773 21.977 1.00 49.68 A ATOM 1932 CB TYR A 255 14.96329.601 20.471 1.00 50.04 A ATOM 1933 CG TYR A 255 15.905 28.596 19.8421.00 50.36 A ATOM 1934 CD1 TYR A 255 15.670 27.228 19.959 1.00 49.51 AATOM 1935 CE1 TYR A 255 16.550 26.296 19.415 1.00 49.32 A ATOM 1936 CD2TYR A 255 17.047 29.012 19.162 1.00 49.32 A ATOM 1937 CE2 TYR A 25517.936 28.088 18.614 1.00 49.59 A ATOM 1938 CZ TYR A 255 17.680 26.73218.746 1.00 48.94 A ATOM 1939 OH TYR A 255 18.557 25.811 18.223 1.0048.35 A ATOM 1940 C TYR A 255 16.545 30.365 22.217 1.00 48.91 A ATOM1941 O TYR A 255 17.471 29.660 22.607 1.00 49.09 A ATOM 1942 N ALA A 25616.659 31.670 21.989 1.00 48.96 A ATOM 1943 CA ALA A 256 17.918 32.39222.145 1.00 49.88 A ATOM 1944 CB ALA A 256 17.756 33.820 21.618 1.0048.87 A ATOM 1945 C ALA A 256 18.500 32.422 23.564 1.00 50.82 A ATOM1946 O ALA A 256 19.716 32.536 23.735 1.00 50.22 A ATOM 1947 N THR A 25717.651 32.328 24.582 1.00 52.17 A ATOM 1948 CA THR A 257 18.152 32.35225.954 1.00 53.69 A ATOM 1949 CB THR A 257 17.347 33.334 26.837 1.0052.46 A ATOM 1950 OG1 THR A 257 15.945 33.092 26.677 1.00 51.52 A ATOM1951 CG2 THR A 257 17.667 34.771 26.462 1.00 52.76 A ATOM 1952 C THR A257 18.145 30.977 26.615 1.00 55.15 A ATOM 1953 O THR A 257 18.70330.804 27.699 1.00 55.75 A ATOM 1954 N LYS A 258 17.528 29.997 25.9591.00 56.37 A ATOM 1955 CA LYS A 258 17.461 28.648 26.514 1.00 56.73 AATOM 1956 CB LYS A 258 16.001 28.247 26.728 1.00 58.42 A ATOM 1957 CGLYS A 258 15.285 29.088 27.773 1.00 59.53 A ATOM 1958 CD LYS A 25813.861 28.613 27.997 1.00 60.71 A ATOM 1959 CE LYS A 258 13.205 29.38729.127 1.00 61.23 A ATOM 1960 NZ LYS A 258 11.785 28.986 29.313 1.0062.40 A ATOM 1961 C LYS A 258 18.160 27.590 25.666 1.00 57.04 A ATOM1962 O LYS A 258 18.072 26.398 25.963 1.00 56.70 A ATOM 1963 N ALA A 25918.860 28.020 24.621 1.00 55.90 A ATOM 1964 CA ALA A 259 19.557 27.08223.743 1.00 55.40 A ATOM 1965 CB ALA A 259 19.969 27.781 22.450 1.0054.73 A ATOM 1966 C ALA A 259 20.780 26.456 24.409 1.00 55.07 A ATOM1967 O ALA A 259 21.536 27.126 25.110 1.00 54.82 A ATOM 1968 N PRO A 26020.988 25.150 24.196 1.00 55.21 A ATOM 1969 CD PRO A 260 20.123 24.18023.501 1.00 54.90 A ATOM 1970 CA PRO A 260 22.139 24.480 24.799 1.0055.01 A ATOM 1971 CB PRO A 260 21.806 23.003 24.615 1.00 55.58 A ATOM1972 CG PRO A 260 21.050 22.999 23.328 1.00 55.22 A ATOM 1973 C PRO A260 23.455 24.860 24.138 1.00 54.81 A ATOM 1974 O PRO A 260 23.52525.030 22.923 1.00 55.25 A ATOM 1975 N TYR A 261 24.493 25.011 24.9521.00 54.39 A ATOM 1976 CA TYR A 261 25.818 25.337 24.452 1.00 55.68 AATOM 1977 CB TYR A 261 25.992 26.853 24.287 1.00 55.85 A ATOM 1978 CGTYR A 261 25.843 27.685 25.542 1.00 55.90 A ATOM 1979 CD1 TYR A 26126.945 28.326 26.108 1.00 55.27 A ATOM 1980 CE1 TYR A 261 26.809 29.13927.232 1.00 55.94 A ATOM 1981 CD2 TYR A 261 24.594 27.873 26.136 1.0055.74 A ATOM 1982 CE2 TYR A 261 24.446 28.682 27.261 1.00 56.36 A ATOM1983 CZ TYR A 261 25.558 29.313 27.803 1.00 56.36 A ATOM 1984 OH TYR A261 25.416 30.118 28.911 1.00 57.53 A ATOM 1985 C TYR A 261 26.85124.761 25.408 1.00 57.21 A ATOM 1986 O TYR A 261 26.639 24.738 26.6231.00 57.77 A ATOM 1987 N ALA A 262 27.958 24.279 24.855 1.00 57.89 AATOM 1988 CA ALA A 262 29.010 23.668 25.655 1.00 58.96 A ATOM 1989 CBALA A 262 29.794 22.674 24.800 1.00 58.74 A ATOM 1990 C ALA A 262 29.96324.668 26.296 1.00 59.61 A ATOM 1991 O ALA A 262 30.337 24.515 27.4601.00 60.65 A ATOM 1992 N TYR A 263 30.352 25.687 25.539 1.00 59.26 AATOM 1993 CA TYR A 263 31.276 26.705 26.027 1.00 58.56 A ATOM 1994 CBTYR A 263 31.688 27.627 24.872 1.00 57.62 A ATOM 1995 CG TYR A 26330.532 28.126 24.031 1.00 55.64 A ATOM 1996 CD1 TYR A 263 29.825 27.26123.193 1.00 55.16 A ATOM 1997 CE1 TYR A 263 28.753 27.712 22.431 1.0053.62 A ATOM 1998 CD2 TYR A 263 30.134 29.460 24.082 1.00 55.16 A ATOM1999 CE2 TYR A 263 29.061 29.922 23.323 1.00 53.97 A ATOM 2000 CZ TYR A263 28.375 29.044 22.503 1.00 53.51 A ATOM 2001 OH TYR A 263 27.29629.494 21.778 1.00 51.12 A ATOM 2002 C TYR A 263 30.762 27.547 27.2021.00 58.82 A ATOM 2003 O TYR A 263 30.686 28.773 27.109 1.00 58.03 AATOM 2004 N VAL A 264 30.419 26.889 28.307 1.00 59.23 A ATOM 2005 CA VALA 264 29.941 27.593 29.496 1.00 59.65 A ATOM 2006 CB VAL A 264 29.57626.607 30.634 1.00 60.55 A ATOM 2007 CG1 VAL A 264 29.253 27.377 31.9111.00 61.12 A ATOM 2008 CG2 VAL A 264 28.384 25.754 30.220 1.00 60.60 AATOM 2009 C VAL A 264 31.034 28.539 29.990 1.00 59.17 A ATOM 2010 O VALA 264 32.211 28.175 30.033 1.00 58.78 A ATOM 2011 N GLY A 265 30.63629.751 30.359 1.00 58.41 A ATOM 2012 CA GLY A 265 31.591 30.739 30.8241.00 57.69 A ATOM 2013 C GLY A 265 31.473 31.966 29.945 1.00 57.54 AATOM 2014 O GLY A 265 31.800 33.086 30.341 1.00 56.90 A ATOM 2015 N GLUA 266 31.009 31.740 28.724 1.00 57.93 A ATOM 2016 CA GLU A 266 30.81138.820 27.777 1.00 58.46 A ATOM 2017 CB GLU A 266 31.588 32.567 26.4901.00 58.78 A ATOM 2018 CG GLU A 266 33.066 32.327 26.702 1.00 59.25 AATOM 2019 CD GLU A 266 33.790 32.041 25.405 1.00 59.38 A ATOM 2020 OE1GLU A 266 34.004 32.986 24.616 1.00 59.35 A ATOM 2021 OE2 GLU A 26634.135 30.865 25.171 1.00 60.57 A ATOM 2022 C GLU A 266 29.326 32.84427.480 1.00 58.93 A ATOM 2023 O GLU A 266 28.578 31.961 27.913 1.0059.06 A ATOM 2024 N LYS A 267 28.893 33.855 26.743 1.00 58.87 A ATOM2025 CA LYS A 267 27.489 33.963 26.403 1.00 58.23 A ATOM 2026 CB LYS A267 27.080 35.438 26.338 1.00 59.85 A ATOM 2027 CG LYS A 267 27.06136.127 27.703 1.00 61.48 A ATOM 2028 CD LYS A 267 28.411 36.008 28.4061.00 62.59 A ATOM 2029 CE LYS A 267 28.296 36.229 29.907 1.00 62.62 AATOM 2030 NZ LYS A 267 29.556 35.847 30.602 1.00 60.90 A ATOM 2031 C LYSA 267 27.256 33.268 25.073 1.00 56.21 A ATOM 2032 O LYS A 267 28.15433.213 24.230 1.00 55.89 A ATOM 2033 N ARG A 268 26.057 32.717 24.9031.00 54.39 A ATOM 2034 CA ARG A 268 25.698 32.023 23.674 1.00 52.55 AATOM 2035 CB ARG A 268 24.194 31.750 23.605 1.00 51.64 A ATOM 2036 CGARG A 268 23.639 30.751 24.585 1.00 52.17 A ATOM 2037 CD ARG A 26822.164 30.533 24.291 1.00 53.30 A ATOM 2038 NE ARG A 268 21.522 29.64125.251 1.00 55.82 A ATOM 2039 CZ ARG A 268 21.311 29.938 26.530 1.0056.60 A ATOM 2040 NH1 ARG A 268 21.689 31.114 27.014 1.00 57.37 A ATOM2041 NH2 ARG A 268 20.722 29.055 27.325 1.00 57.84 A ATOM 2042 C ARG A268 26.053 32.847 22.454 1.00 50.70 A ATOM 2043 O ARG A 268 25.85634.065 22.435 1.00 51.42 A ATOM 2044 N THR A 269 26.585 32.176 21.4411.00 50.18 A ATOM 2045 CA THR A 269 26.902 32.828 20.183 1.00 47.44 AATOM 2046 CB THR A 269 28.116 32.200 19.498 1.00 48.13 A ATOM 2047 OG1THR A 269 29.289 32.466 20.277 1.00 48.94 A ATOM 2048 CG2 THR A 26928.297 32.784 18.096 1.00 47.82 A ATOM 2049 C THR A 269 25.656 32.57219.343 1.00 46.15 A ATOM 2050 O THR A 269 25.227 31.430 19.188 1.0046.11 A ATOM 2051 N LEU A 270 25.063 33.641 18.830 1.00 44.47 A ATOM2052 CA LEU A 270 23.853 33.532 18.031 1.00 43.06 A ATOM 2053 CB LEU A270 22.713 34.265 18.740 1.00 43.60 A ATOM 2054 CG LEU A 270 22.46833.872 20.200 1.00 44.62 A ATOM 2055 CD1 LEU A 270 21.764 35.002 20.9271.00 44.69 A ATOM 2056 CD2 LEU A 270 21.646 32.597 20.261 1.00 44.54 AATOM 2057 C LEU A 270 24.054 34.133 16.642 1.00 40.92 A ATOM 2058 O LEUA 270 24.858 35.046 16.462 1.00 42.68 A ATOM 2059 N VAL A 271 23.33233.603 15.663 1.00 39.89 A ATOM 2060 CA VAL A 271 23.394 34.120 14.2981.00 38.73 A ATOM 2061 CB VAL A 271 24.243 33.236 13.371 1.00 39.03 AATOM 2062 CG1 VAL A 271 24.009 33.628 11.911 1.00 37.91 A ATOM 2063 CG2VAL A 271 25.703 33.419 13.699 1.00 37.99 A ATOM 2064 C VAL A 271 21.97234.181 13.775 1.00 37.49 A ATOM 2065 O VAL A 271 21.330 33.154 13.5681.00 36.59 A ATOM 2066 N TYR A 272 21.475 35.398 13.602 1.00 37.97 AATOM 2067 CA TYR A 272 20.121 35.605 13.113 1.00 36.60 A ATOM 2068 CBTYR A 272 19.528 36.903 13.659 1.00 37.13 A ATOM 2069 CG TYR A 27219.312 36.932 15.152 1.00 40.37 A ATOM 2070 CD1 TYR A 272 20.387 37.05616.033 1.00 40.17 A ATOM 2071 CE1 TYR A 272 20.178 37.136 17.413 1.0042.29 A ATOM 2072 CD2 TYR A 272 18.021 36.879 15.685 1.00 41.07 A ATOM2073 CE2 TYR A 272 17.805 36.956 17.056 1.00 41.54 A ATOM 2074 CZ TYR A272 18.883 37.086 17.913 1.00 41.60 A ATOM 2075 OH TYR A 272 18.65737.172 19.266 1.00 43.79 A ATOM 2076 C TYR A 272 20.109 35.686 11.6031.00 35.69 A ATOM 2077 O TYR A 272 21.119 35.983 10.977 1.00 34.57 AATOM 2078 N GLY A 273 18.940 35.444 11.031 1.00 34.52 A ATOM 2079 CA GLYA 273 18.815 35.508 9.596 1.00 34.19 A ATOM 2080 C GLY A 273 17.48736.075 9.159 1.00 32.96 A ATOM 2081 O GLY A 273 16.435 35.786 9.747 1.0033.22 A ATOM 2082 N LEU A 274 17.548 36.915 8.136 1.00 31.58 A ATOM 2083CA LEU A 274 16.360 37.508 7.557 1.00 30.07 A ATOM 2084 CB LEU A 27416.417 39.029 7.606 1.00 30.58 A ATOM 2085 CG LEU A 274 15.329 39.7126.766 1.00 32.48 A ATOM 2086 CD1 LEU A 274 13.961 39.283 7.256 1.0032.49 A ATOM 2087 CD2 LEU A 274 15.470 41.216 6.855 1.00 31.20 A ATOM2088 C LEU A 274 16.403 37.044 6.119 1.00 29.70 A ATOM 2089 O LEU A 27417.318 37.399 5.377 1.00 27.86 A ATOM 2090 N THR A 275 15.429 36.2315.737 1.00 28.56 A ATOM 2091 CA THR A 275 15.384 35.704 4.390 1.00 30.21A ATOM 2092 CB THR A 275 15.249 34.174 4.397 1.00 30.89 A ATOM 2093 OG1THR A 275 16.243 33.611 5.263 1.00 31.27 A ATOM 2094 CG2 THR A 27515.466 33.617 2.985 1.00 32.12 A ATOM 2095 C THR A 275 14.228 36.2963.613 1.00 31.45 A ATOM 2096 O THR A 275 13.095 36.367 4.106 1.00 30.04A ATOM 2097 N LEU A 276 14.534 36.707 2.389 1.00 30.98 A ATOM 2098 CALEU A 276 13.560 37.306 1.502 1.00 32.05 A ATOM 2099 CB LEU A 276 14.02938.692 1.063 1.00 32.02 A ATOM 2100 CG LEU A 276 14.463 39.658 2.1611.00 31.99 A ATOM 2101 CD1 LEU A 276 15.012 40.926 1.528 1.00 33.02 AATOM 2102 CD2 LEU A 276 13.287 39.968 3.067 1.00 31.46 A ATOM 2103 C LEUA 276 13.377 36.442 0.269 1.00 32.27 A ATOM 2104 O LEU A 276 14.33136.202 −0.471 1.00 31.75 A ATOM 2105 N SER A 277 12.151 35.967 0.0651.00 33.88 A ATOM 2106 CA SER A 277 11.830 35.164 −1.101 1.00 34.99 AATOM 2107 CB SER A 277 11.166 33.848 −0.689 1.00 35.49 A ATOM 2108 OGSER A 277 9.833 34.061 −0.264 1.00 37.40 A ATOM 2109 C SER A 277 10.87336.020 −1.931 1.00 36.36 A ATOM 2110 O SER A 277 10.451 37.090 −1.4931.00 34.72 A ATOM 2111 N PRO A 278 10.514 35.563 −3.139 1.00 39.44 AATOM 2112 CD PRO A 278 10.974 34.375 −3.875 1.00 39.05 A ATOM 2113 CAPRO A 278 9.606 36.369 −3.957 1.00 40.64 A ATOM 2114 CB PRO A 278 9.43335.521 −5.214 1.00 40.18 A ATOM 2115 CG PRO A 278 10.752 34.792 −5.3061.00 40.69 A ATOM 2116 C PRO A 278 8.272 36.695 −3.300 1.00 42.09 A ATOM2117 O PRO A 278 7.765 37.807 −3.428 1.00 43.87 A ATOM 2118 N LYS A 2797.715 35.740 −2.570 1.00 44.32 A ATOM 2119 CA LYS A 279 6.414 35.959−1.964 1.00 45.01 A ATOM 2120 CB LYS A 279 5.533 34.738 −2.224 1.0046.55 A ATOM 2121 CG LYS A 279 5.493 34.307 −3.693 1.00 48.65 A ATOM2122 CD LYS A 279 4.868 35.368 −4.594 1.00 49.82 A ATOM 2123 CE LYS A279 4.878 34.913 −6.054 1.00 50.84 A ATOM 2124 NZ LYS A 279 4.280 35.916−6.979 1.00 51.34 A ATOM 2125 C LYS A 279 6.380 36.303 −0.483 1.00 44.70A ATOM 2126 O LYS A 279 5.385 36.834 0.001 1.00 45.30 A ATOM 2127 N GLUA 280 7.457 36.029 0.240 1.00 43.74 A ATOM 2128 CA GLU A 280 7.46436.308 1.670 1.00 42.41 A ATOM 2129 CB GLU A 280 6.956 35.086 2.439 1.0045.17 A ATOM 2130 CG GLU A 280 7.607 33.770 1.997 1.00 48.54 A ATOM 2131CD GLU A 280 7.531 32.665 3.051 1.00 51.12 A ATOM 2132 OE1 GLU A 2806.653 32.737 3.940 1.00 53.02 A ATOM 2133 OE2 GLU A 280 8.346 31.7172.982 1.00 50.28 A ATOM 2134 C GLU A 280 8.816 36.688 2.244 1.00 40.99 AATOM 2135 O GLU A 280 9.796 36.885 1.527 1.00 37.74 A ATOM 2136 N GLU A281 8.828 36.773 3.569 1.00 40.26 A ATOM 2137 CA GLU A 281 10.003 37.0794.363 1.00 39.86 A ATOM 2138 CB GLU A 281 10.008 38.562 4.754 1.00 40.81A ATOM 2139 CG GLU A 281 10.698 38.866 6.077 1.00 41.06 A ATOM 2140 CDGLU A 281 10.540 40.316 6.517 1.00 43.02 A ATOM 2141 OE1 GLU A 28110.990 41.219 5.784 1.00 42.38 A ATOM 2142 OE2 GLU A 281 9.967 40.5587.603 1.00 42.59 A ATOM 2143 C GLU A 281 9.874 36.224 5.621 1.00 39.25 AATOM 2144 O GLU A 281 8.813 36.204 6.242 1.00 39.38 A ATOM 2145 N TYR A282 10.929 35.501 5.984 1.00 36.51 A ATOM 2146 CA TYR A 282 10.89534.703 7.200 1.00 35.17 A ATOM 2147 CB TYR A 282 10.575 33.231 6.9091.00 34.96 A ATOM 2148 CG TYR A 282 11.546 32.493 6.022 1.00 35.52 AATOM 2149 CD1 TYR A 282 12.574 31.728 6.569 1.00 35.03 A ATOM 2150 CE1TYR A 282 13.446 31.001 5.758 1.00 36.48 A ATOM 2151 CD2 TYR A 28211.411 32.521 4.631 1.00 34.37 A ATOM 2152 CE2 TYR A 282 12.279 31.7963.809 1.00 34.23 A ATOM 2153 CZ TYR A 282 13.291 31.040 4.380 1.00 34.88A ATOM 2154 OH TYR A 282 14.143 30.310 3.584 1.00 37.85 A ATOM 2155 CTYR A 282 12.205 34.871 7.956 1.00 34.80 A ATOM 2156 O TYR A 282 13.18235.398 7.415 1.00 32.60 A ATOM 2157 N TYR A 283 12.216 34.423 9.206 1.0034.19 A ATOM 2158 CA TYR A 283 13.369 34.598 10.080 1.00 33.80 A ATOM2159 CB TYR A 283 12.910 35.412 11.288 1.00 35.23 A ATOM 2160 CG TYR A283 11.860 36.424 10.889 1.00 36.91 A ATOM 2161 CD1 TYR A 283 12.19337.541 10.117 1.00 37.31 A ATOM 2162 CE1 TYR A 283 11.206 38.407 9.6291.00 37.64 A ATOM 2163 CD2 TYR A 283 10.512 36.203 11.175 1.00 38.08 AATOM 2164 CE2 TYR A 283 9.521 37.059 10.695 1.00 38.79 A ATOM 2165 CZTYR A 283 9.875 38.154 9.923 1.00 39.16 A ATOM 2166 OH TYR A 283 8.89038.986 9.446 1.00 39.66 A ATOM 2167 C TYR A 283 14.041 33.302 10.5191.00 33.97 A ATOM 2168 O TYR A 283 13.404 32.258 10.627 1.00 34.03 AATOM 2169 N LYS A 284 15.342 33.375 10.768 1.00 32.82 A ATOM 2170 CA LYSA 284 16.086 32.191 11.170 1.00 33.38 A ATOM 2171 CB LYS A 284 16.89331.668 9.974 1.00 34.19 A ATOM 2172 CG LYS A 284 15.989 31.260 8.8091.00 36.80 A ATOM 2173 CD LYS A 284 16.733 30.629 7.633 1.00 39.12 AATOM 2174 CE LYS A 284 17.536 31.643 6.843 1.00 40.11 A ATOM 2175 NZ LYSA 284 18.047 31.069 5.548 1.00 34.37 A ATOM 2176 C LYS A 284 16.98832.481 12.362 1.00 32.66 A ATOM 2177 O LYS A 284 17.392 33.624 12.5831.00 33.84 A ATOM 2178 N LEU A 285 17.283 31.448 13.143 1.00 33.01 AATOM 2179 CA LEU A 285 18.141 31.621 14.308 1.00 33.94 A ATOM 2180 CBLEU A 285 17.309 32.021 15.531 1.00 34.70 A ATOM 2181 CG LEU A 28518.063 32.102 16.862 1.00 36.10 A ATOM 2182 CD1 LEU A 285 19.214 33.08516.752 1.00 35.16 A ATOM 2183 CD2 LEU A 285 17.097 32.525 17.965 1.0036.19 A ATOM 2184 C LEU A 285 18.950 30.377 14.633 1.00 33.22 A ATOM2185 O LEU A 285 18.401 29.311 14.918 1.00 34.61 A ATOM 2186 N GLY A 28620.265 30.527 14.582 1.00 33.99 A ATOM 2187 CA GLY A 286 21.142 29.42214.893 1.00 34.28 A ATOM 2188 C GLY A 286 21.824 29.682 16.218 1.0034.79 A ATOM 2189 O GLY A 286 22.402 30.745 16.418 1.00 35.42 A ATOM2190 N ALA A 287 21.728 28.726 17.135 1.00 36.63 A ATOM 2191 CA ALA A287 22.369 28.848 18.439 1.00 37.80 A ATOM 2192 CB ALA A 287 21.38428.517 19.546 1.00 37.09 A ATOM 2193 C ALA A 287 23.530 27.865 18.4541.00 38.30 A ATOM 2194 O ALA A 287 23.326 26.648 18.412 1.00 38.88 AATOM 2195 N TYR A 288 24.746 28.397 18.497 1.00 40.30 A ATOM 2196 CA TYRA 288 25.936 27.569 18.505 1.00 42.62 A ATOM 2197 CB TYR A 288 27.18928.434 18.343 1.00 43.09 A ATOM 2198 CG TYR A 288 27.362 29.009 16.9551.00 43.83 A ATOM 2199 CD1 TYR A 288 26.405 29.865 16.409 1.00 45.17 AATOM 2200 CE1 TYR A 288 26.545 30.373 15.119 1.00 44.97 A ATOM 2201 CD2TYR A 288 28.469 28.679 16.175 1.00 43.95 A ATOM 2202 CE2 TYR A 28828.621 29.182 14.885 1.00 43.70 A ATOM 2203 CZ TYR A 288 27.654 30.02714.365 1.00 45.56 A ATOM 2204 OH TYR A 288 27.789 30.525 13.090 1.0046.87 A ATOM 2205 C TYR A 288 26.059 26.734 19.768 1.00 45.18 A ATOM2206 O TYR A 288 25.919 27.240 20.882 1.00 44.32 A ATOM 2207 N TYR A 28926.305 25.443 19.573 1.00 46.25 A ATOM 2208 CA TYR A 289 26.496 24.51920.674 1.00 47.59 A ATOM 2209 CB TYR A 289 25.997 23.126 20.305 1.0048.44 A ATOM 2210 CG TYR A 289 26.224 22.105 21.393 1.00 51.30 A ATOM2211 CD1 TYR A 289 25.700 22.294 22.674 1.00 51.82 A ATOM 2212 CE1 TYR A289 25.937 21.378 23.688 1.00 53.60 A ATOM 2213 CD2 TYR A 289 26.98820.965 21.155 1.00 50.81 A ATOM 2214 CE2 TYR A 289 27.230 20.041 22.1621.00 53.45 A ATOM 2215 CZ TYR A 289 26.702 20.255 23.426 1.00 53.78 AATOM 2216 OH TYR A 289 26.946 19.346 24.428 1.00 55.63 A ATOM 2217 C TYRA 289 27.998 24.496 20.912 1.00 47.53 A ATOM 2218 O TYR A 289 28.45824.476 22.053 1.00 48.27 A ATOM 2219 N HIS A 290 28.755 24.505 19.8191.00 46.65 A ATOM 2220 CA HIS A 290 30.214 24.524 19.871 1.00 45.99 AATOM 2221 CB HIS A 290 30.824 23.263 19.246 1.00 47.09 A ATOM 2222 CGHIS A 290 30.663 22.026 20.070 1.00 47.66 A ATOM 2223 CD2 HIS A 29030.004 21.803 21.231 1.00 48.23 A ATOM 2224 ND1 HIS A 290 31.203 20.81519.697 1.00 48.48 A ATOM 2225 CE1 HIS A 290 30.880 19.897 20.592 1.0048.01 A ATOM 2226 NE2 HIS A 290 30.152 20.471 21.532 1.00 47.65 A ATOM2227 C HIS A 290 30.695 25.715 19.061 1.00 46.29 A ATOM 2228 O HIS A 29030.152 26.000 17.986 1.00 45.19 A ATOM 2229 N ILE A 291 31.708 26.40719.573 1.00 44.93 A ATOM 2230 CA ILE A 291 32.282 27.548 18.875 1.0045.16 A ATOM 2231 CB ILE A 291 31.889 28.907 19.524 1.00 45.11 A ATOM2232 CG2 ILE A 291 30.417 29.188 19.301 1.00 43.33 A ATOM 2233 CG1 ILE A291 32.243 28.899 21.019 1.00 45.59 A ATOM 2234 CD1 ILE A 291 32.18630.259 21.675 1.00 45.30 A ATOM 2235 C ILE A 291 33.798 27.454 18.8791.00 45.61 A ATOM 2236 O ILE A 291 34.387 26.651 19.602 1.00 47.34 AATOM 2237 N THR A 292 34.424 28.291 18.065 1.00 47.17 A ATOM 2238 CA THRA 292 35.875 28.341 17.967 1.00 47.28 A ATOM 2239 CB THR A 292 36.37127.755 16.639 1.00 47.48 A ATOM 2240 OG1 THR A 292 35.993 28.517 15.5551.00 44.68 A ATOM 2241 CG2 THR A 292 35.775 26.374 16.426 1.00 46.70 AATOM 2242 C THR A 292 36.212 29.814 18.001 1.00 47.83 A ATOM 2243 O THRA 292 35.307 30.644 18.064 1.00 48.07 A ATOM 2244 N ASP A 293 37.49730.149 17.954 1.00 48.59 A ATOM 2245 CA ASP A 293 37.883 31.551 17.9821.00 48.64 A ATOM 2246 CB ASP A 293 39.408 31.705 17.988 1.00 49.60 AATOM 2247 CG ASP A 293 40.045 31.150 19.249 1.00 50.84 A ATOM 2248 OD1ASP A 293 39.394 31.201 20.317 1.00 52.13 A ATOM 2249 OD2 ASP A 29341.200 30.673 19.176 1.00 51.40 A ATOM 2250 C ASP A 293 37.299 32.28416.784 1.00 48.56 A ATOM 2251 O ASP A 293 37.058 33.488 16.851 1.0049.15 A ATOM 2252 N VAL A 294 37.070 31.561 15.689 1.00 47.81 A ATOM2253 CA VAL A 294 36.512 32.179 14.487 1.00 46.87 A ATOM 2254 CB VAL A294 36.140 31.120 13.418 1.00 47.55 A ATOM 2255 CG1 VAL A 294 35.34631.772 12.291 1.00 47.30 A ATOM 2256 CG2 VAL A 294 37.409 30.490 12.8471.00 47.74 A ATOM 2257 C VAL A 294 35.272 32.985 14.843 1.00 45.87 AATOM 2258 O VAL A 294 35.123 34.130 14.416 1.00 46.94 A ATOM 2259 N GLNA 295 34.389 32.380 15.629 1.00 45.48 A ATOM 2260 CA GLN A 295 33.16233.035 16.061 1.00 45.44 A ATOM 2261 CB GLN A 295 32.190 32.002 16.6471.00 45.04 A ATOM 2262 CG GLN A 295 31.550 31.080 15.603 1.00 44.79 AATOM 2263 CD GLN A 295 32.555 30.174 14.906 1.00 45.62 A ATOM 2264 OE1GLN A 295 32.408 29.860 13.720 1.00 44.71 A ATOM 2265 NE2 GLN A 29533.576 29.741 15.642 1.00 42.80 A ATOM 2266 C GLN A 295 33.472 34.11817.096 1.00 45.66 A ATOM 2267 O GLN A 295 32.840 35.178 17.109 1.0044.55 A ATOM 2268 N ARG A 296 34.449 33.850 17.960 1.00 45.23 A ATOM2269 CA ARG A 296 34.842 34.819 18.982 1.00 45.03 A ATOM 2270 CB ARG A296 35.964 34.255 19.862 1.00 44.33 A ATOM 2271 CG ARG A 296 35.59232.952 20.540 1.00 44.26 A ATOM 2272 CD ARG A 296 36.126 32.857 21.9701.00 45.12 A ATOM 2273 NE ARG A 296 35.665 31.624 22.597 1.00 44.32 AATOM 2274 CZ ARG A 296 36.124 30.419 22.280 1.00 44.21 A ATOM 2275 NH1ARG A 296 37.069 30.292 21.357 1.00 44.51 A ATOM 2276 NH2 ARG A 29635.613 29.341 22.860 1.00 45.05 A ATOM 2277 C ARG A 296 35.317 36.09118.297 1.00 44.39 A ATOM 2278 O ARG A 296 35.067 37.202 18.766 1.0044.29 A ATOM 2279 N GLY A 297 36.004 35.917 17.175 1.00 43.83 A ATOM2280 CA GLY A 297 36.494 37.061 16.436 1.00 44.22 A ATOM 2281 C GLY A297 35.372 37.832 15.766 1.00 45.50 A ATOM 2282 O GLY A 297 35.37039.064 15.771 1.00 46.22 A ATOM 2283 N LEU A 298 34.417 37.108 15.1881.00 45.75 A ATOM 2284 CA LEU A 298 33.292 37.732 14.498 1.00 46.53 AATOM 2285 CB LEU A 298 32.402 36.663 13.848 1.00 45.17 A ATOM 2286 CGLEU A 298 33.021 35.931 12.650 1.00 45.06 A ATOM 2287 CD1 LEU A 29832.084 34.824 12.164 1.00 44.48 A ATOM 2288 CD2 LEU A 298 33.307 36.93211.537 1.00 45.31 A ATOM 2289 C LEU A 298 32.462 38.602 15.433 1.0047.62 A ATOM 2290 O LEU A 298 31.945 39.646 15.028 1.00 47.53 A ATOM2291 N LEU A 299 32.340 38.171 16.685 1.00 48.72 A ATOM 2292 CA LEU A299 31.580 38.924 17.667 1.00 50.18 A ATOM 2293 CB LEU A 299 31.39038.090 18.930 1.00 49.60 A ATOM 2294 CG LEU A 299 30.350 36.975 18.8071.00 48.08 A ATOM 2295 CD1 LEU A 299 30.486 36.019 19.974 1.00 47.37 AATOM 2296 CD2 LEU A 299 28.953 37.573 18.765 1.00 48.29 A ATOM 2297 CLEU A 299 32.240 40.260 18.004 1.00 51.67 A ATOM 2298 O LEU A 299 31.55041.240 18.282 1.00 51.55 A ATOM 2299 N LYS A 300 33.572 40.303 17.9701.00 53.35 A ATOM 2300 CA LYS A 300 34.304 41.537 18.263 1.00 53.76 AATOM 2301 CB LYS A 300 35.794 41.265 18.493 1.00 54.63 A ATOM 2302 CGLYS A 300 36.147 40.439 19.703 1.00 56.78 A ATOM 2303 CD LYS A 30037.663 40.361 19.835 1.00 58.59 A ATOM 2304 CE LYS A 300 38.091 39.47320.991 1.00 60.33 A ATOM 2305 NZ LYS A 300 39.558 39.580 21.244 1.0061.89 A ATOM 2306 C LYS A 300 34.192 42.492 17.086 1.00 53.47 A ATOM2307 O LYS A 300 34.072 43.705 17.259 1.00 54.79 A ATOM 2308 N ALA A 30134.241 41.933 15.885 1.00 51.85 A ATOM 2309 CA ALA A 301 34.172 42.72914.669 1.00 50.07 A ATOM 2310 CB ALA A 301 34.698 41.913 13.495 1.0050.60 A ATOM 2311 C ALA A 301 32.791 43.272 14.332 1.00 49.04 A ATOM2312 O ALA A 301 32.672 44.404 13.855 1.00 48.99 A ATOM 2313 N PHE A 30231.750 42.479 14.583 1.00 47.91 A ATOM 2314 CA PHE A 302 30.391 42.89514.244 1.00 46.87 A ATOM 2315 CB PHE A 302 29.775 41.920 13.229 1.0045.69 A ATOM 2316 CG PHE A 302 30.542 41.819 11.943 1.00 41.83 A ATOM2317 CD1 PHE A 302 31.567 40.890 11.801 1.00 41.69 A ATOM 2318 CD2 PHE A302 30.256 42.672 10.881 1.00 42.51 A ATOM 2319 CE1 PHE A 302 32.29840.813 10.620 1.00 40.43 A ATOM 2320 CE2 PHE A 302 30.982 42.603 9.6931.00 40.70 A ATOM 2321 CZ PHE A 302 32.004 41.675 9.562 1.00 41.10 AATOM 2322 C PHE A 302 29.418 43.061 15.398 1.00 47.58 A ATOM 2323 O PHEA 302 28.303 43.536 15.198 1.00 47.34 A ATOM 2324 N ASP A 303 29.81742.668 16.600 1.00 48.39 A ATOM 2325 CA ASP A 303 28.919 42.807 17.7381.00 49.75 A ATOM 2326 CB ASP A 303 28.481 41.423 18.224 1.00 48.37 AATOM 2327 CG ASP A 303 27.185 41.462 18.998 1.00 47.54 A ATOM 2328 OD1ASP A 303 26.215 42.066 18.497 1.00 47.88 A ATOM 2329 OD2 ASP A 30327.126 40.879 20.100 1.00 49.46 A ATOM 2330 C ASP A 303 29.581 43.58918.874 1.00 50.31 A ATOM 2331 O ASP A 303 29.661 43.050 19.995 1.0051.55 A ATOM 2332 OXT ASP A 303 30.012 44.737 18.627 1.00 51.61 A ATOM2333 MG + 2 MG2 A 1 19.221 25.308 7.555 1.00 43.67 M ATOM 2334 PA GSP A1 19.534 27.534 3.619 1.00 40.67 G ATOM 2335 O1A GSP A 1 19.609 28.2042.288 1.00 39.03 G ATOM 2336 O2A GSP A 1 20.362 26.998 3.674 1.00 39.43G ATOM 2337 O3A GSP A 1 18.121 27.257 4.002 1.00 42.12 G ATOM 2338 O1BGSP A 1 20.124 28.585 4.679 1.00 40.27 G ATOM 2339 PB GSP A 1 20.52928.366 6.198 1.00 40.45 G ATOM 2340 O2B GSP A 1 20.225 29.576 7.030 1.0040.45 G ATOM 2341 O3B GSP A 1 19.816 27.203 6.821 1.00 40.17 G ATOM 2342S1 GSP A 1 22.118 28.106 6.368 1.00 46.86 G ATOM 2343 C1 GSP A 1 22.78528.278 7.611 1.00 48.16 G ATOM 2344 C2 GSP A 1 23.948 27.320 7.744 1.0048.30 G ATOM 2345 C3 GSP A 1 25.237 27.674 7.899 1.00 48.85 G ATOM 2346C10 GSP A 1 25.862 28.840 7.172 1.00 48.34 G ATOM 2347 C4 GSP A 1 26.16526.878 8.794 1.00 49.42 G ATOM 2348 C5 GSP A 1 27.559 27.525 8.868 1.0051.23 G ATOM 2349 C6 GSP A 1 28.644 26.503 9.103 1.00 53.24 G ATOM 2350C7 GSP A 1 29.414 26.374 10.200 1.00 53.60 G ATOM 2351 C9 GSP A 1 30.50527.374 10.507 1.00 54.65 G ATOM 2352 C8 GSP A 1 29.296 25.257 11.2111.00 55.35 G ATOM 2353 C1 DH2 A 1 26.153 31.881 8.710 1.00 65.18 D ATOM2354 C2 DH2 A 1 24.959 31.390 9.301 1.00 65.39 D ATOM 2355 C3 DH2 A 125.028 30.627 10.451 1.00 66.41 D ATOM 2356 C4 DH2 A 1 26.279 30.32411.055 1.00 66.26 D ATOM 2357 C5 DH2 A 1 27.435 30.796 10.488 1.00 66.06D ATOM 2358 C6 DH2 A 1 28.585 32.099 8.675 1.00 64.51 D ATOM 2359 C7 DH2A 1 27.251 33.138 6.948 1.00 63.33 D ATOM 2360 C8 DH2 A 1 26.099 32.6637.525 1.00 63.43 D ATOM 2361 O1 DH2 A 1 29.857 31.880 9.155 1.00 65.00 DATOM 2362 O2 DH2 A 1 23.839 30.179 10.983 1.00 66.99 D ATOM 2363 CP DH2A 1 28.509 32.861 7.519 1.00 64.14 D ATOM 2364 C10 DH2 A 1 27.409 31.5849.306 1.00 65.41 D ATOM 2365 N NO3 A 1 42.060 34.777 −5.122 1.00 69.01 NATOM 2366 O1 NO3 A 1 41.197 34.277 −5.872 1.00 69.03 N ATOM 2367 O2 NO3A 1 41.725 35.701 −4.452 1.00 68.04 N ATOM 2368 O3 NO3 A 1 43.135 34.230−5.061 1.00 68.81 N ATOM 2369 OH2 TIP A 1 30.490 35.181 26.300 1.0065.42 W ATOM 2370 OH2 TIP A 2 36.852 21.356 11.220 1.00 62.71 W ATOM2371 OH2 TIP A 3 13.275 6.277 5.899 1.00 52.03 W ATOM 2372 OH2 TIP A 439.421 26.056 19.137 1.00 55.68 W ATOM 2373 OH2 TIP A 5 29.221 47.05219.171 1.00 51.85 W ATOM 2374 OH2 TIP A 6 25.560 30.322 10.756 1.0097.83 W ATOM 2375 OH2 TIP A 7 23.552 52.697 −2.797 1.00 48.74 W ATOM2376 OH2 TIP A 8 15.783 28.067 4.579 1.00 42.75 W ATOM 2377 OH2 TIP A 935.855 25.568 −13.555 1.00 79.37 W ATOM 2378 OH2 TIP A 10 36.538 42.420−0.112 1.00 49.25 W ATOM 2379 OH2 TIP A 11 23.745 35.145 24.150 1.0053.80 W ATOM 2380 OH2 TIP A 12 19.673 51.064 6.990 1.00 47.69 W ATOM2381 OH2 TIP A 13 27.368 30.522 28.673 1.00 65.03 W ATOM 2382 OH2 TIP A14 32.186 14.701 −3.539 1.00 56.64 W ATOM 2383 OH2 TIP A 15 24.46014.475 −13.361 1.00 59.07 W ATOM 2384 OH2 TIP A 16 40.585 17.057 1.4711.00 59.15 W ATOM 2385 OH2 TIP A 17 17.913 13.365 −5.351 1.00 45.07 WATOM 2386 OH2 TIP A 18 15.723 27.642 7.214 1.00 47.54 W ATOM 2387 OH2TIP A 19 22.263 7.047 −6.225 1.00 59.29 W ATOM 2388 OH2 TIP A 20 19.96929.141 −17.580 1.00 67.73 W ATOM 2389 OH2 TIP A 21 21.634 48.493 12.1711.00 47.20 W ATOM 2390 OH2 TIP A 22 18.106 22.881 −16.847 1.00 50.83 WATOM 2391 OH2 TIP A 23 29.495 40.536 21.854 1.00 60.58 W ATOM 2392 OH2TIP A 24 36.302 27.268 13.132 1.00 55.92 W ATOM 2393 OH2 TIP A 25 29.73235.121 4.882 1.00 50.52 W ATOM 2394 OH2 TIP A 26 45.475 29.946 0.2401.00 44.34 W ATOM 2395 OH2 TIP A 27 15.795 10.968 19.889 1.00 77.54 WATOM 2396 OH2 TIP A 28 14.431 30.709 −8.734 1.00 39.58 W ATOM 2397 OH2TIP A 29 13.313 28.671 −1.901 1.00 44.66 W ATOM 2398 OH2 TIP A 30 27.44052.388 8.029 1.00 60.67 W ATOM 2399 OH2 TIP A 31 42.559 22.415 4.0641.00 53.62 W ATOM 2400 OH2 TIP A 32 20.941 43.226 24.809 1.00 55.10 WATOM 2401 OH2 TIP A 33 33.359 44.497 −2.388 1.00 58.45 W ATOM 2402 OH2TIP A 34 31.147 53.165 −0.332 1.00 46.23 W ATOM 2403 OH2 TIP A 35 12.35625.536 1.697 1.00 64.15 W ATOM 2404 OH2 TIP A 36 35.138 10.636 12.6521.00 61.46 W ATOM 2405 OH2 TIP A 37 15.020 50.521 −4.580 1.00 40.30 WATOM 2406 OH2 TIP A 38 9.781 27.922 21.238 1.00 68.26 W ATOM 2407 OH2TIP A 39 41.691 22.533 −2.565 1.00 54.13 W ATOM 2408 OH2 TIP A 40 20.66645.859 15.289 1.00 70.16 W ATOM 2409 OH2 TIP A 41 34.363 37.353 21.3741.00 53.19 W ATOM 2410 OH2 TIP A 42 26.734 5.871 −6.191 1.00 45.50 WATOM 2411 OH2 TIP A 43 10.365 49.794 12.437 1.00 50.70 W ATOM 2412 OH2TIP A 44 34.543 29.704 9.768 1.00 55.92 W ATOM 2413 OH2 TIP A 45 40.37631.076 6.686 1.00 58.22 W ATOM 2414 OH2 TIP A 46 34.553 5.913 23.8581.00 71.52 W ATOM 2415 OH2 TIP A 47 14.200 27.005 5.946 1.00 53.11 WATOM 2416 OH2 TIP A 48 42.976 30.424 −5.522 1.00 44.67 W ATOM 2417 OH2TIP A 49 31.686 41.102 21.600 1.00 49.60 W ATOM 2418 OH2 TIP A 50 37.98529.637 6.203 1.00 60.32 W ATOM 2419 OH2 TIP A 51 16.521 36.553 −8.8981.00 49.77 W ATOM 2420 OH2 TIP A 52 31.859 17.997 −13.734 1.00 53.29 WATOM 2421 OH2 TIP A 53 27.801 29.505 30.891 1.00 49.38 W ATOM 2422 OH2TIP A 54 4.648 43.961 2.873 1.00 57.90 W ATOM 2423 OH2 TIP A 55 18.09343.972 14.582 1.00 66.29 W ATOM 2424 OH2 TIP A 56 32.762 25.967 29.9111.00 57.94 W ATOM 2425 OH2 TIP A 57 6.942 34.079 7.083 1.00 48.37 W ATOM2426 OH2 TIP A 58 43.391 12.772 6.133 1.00 61.46 W ATOM 2427 OH2 TIP A59 8.265 49.640 −10.664 1.00 59.37 W ATOM 2428 OH2 TIP A 60 6.971 18.92310.395 1.00 62.11 W ATOM 2429 OH2 TIP A 61 12.616 28.504 1.451 1.0057.01 W ATOM 2430 OH2 TIP A 62 25.673 48.908 −3.608 1.00 48.26 W ATOM2431 OH2 TIP A 63 11.864 30.819 −2.529 1.00 57.25 W ATOM 2432 OH2 TIP A64 23.756 43.583 −16.811 1.00 53.58 W ATOM 2433 OH2 TIP A 65 14.20719.466 0.982 1.00 52.77 W ATOM 2434 OH2 TIP A 66 2.064 35.531 −8.9841.00 67.76 W ATOM 2435 OH2 TIP A 67 27.154 45.880 16.535 1.00 61.40 WATOM 2436 OH2 TIP A 68 29.710 31.367 12.036 1.00 54.10 W ATOM 2437 OH2TIP A 69 37.562 26.637 4.885 1.00 84.16 W ATOM 2438 OH2 TIP A 70 16.12118.617 −9.991 1.00 46.09 W ATOM 2439 OH2 TIP A 1 28.748 17.651 −3.9731.00 43.52 SS ATOM 2440 OH2 TIP A 2 40.158 21.998 7.668 1.00 43.52 SSATOM 2441 OH2 TIP A 3 20.062 48.816 5.351 1.00 43.52 SS ATOM 2442 OH2TIP A 4 24.944 5.885 −0.742 1.00 43.52 SS ATOM 2443 OH2 TIP A 5 30.68825.005 −4.289 1.00 4352 SS ATOM 2444 OH2 TIP A 6 19.073 30.215 −9.7761.00 43.52 SS ATOM 2445 OH2 TIP A 7 18.367 30.661 2.655 1.00 43.52 SSATOM 2446 OH2 TIP A 8 19.367 32.272 0.565 1.00 43.52 SS ATOM 2447 OH2TIP A 9 29.275 23.177 −0.801 1.00 43.52 SS ATOM 2448 OH2 TIP A 10 30.92616.038 −5.142 1.00 43.52 SS ATOM 2449 OH2 TIP A 11 34.325 28.514 11.4491.00 43.52 SS ATOM 2450 OH2 TIP A 12 21.383 27.492 −13.294 1.00 43.52 SSATOM 2451 OH2 TIP A 13 22.108 28.693 11.655 1.00 43.52 SS ATOM 2452 OH2TIP A 14 18.252 23.094 −9.262 1.00 43.52 SS ATOM 2453 OH2 TIP A 1518.047 20.583 −11.156 1.00 43.52 SS ATOM 2454 OH2 TIP A 16 17.057 52.115−4.118 1.00 43.52 SS ATOM 2455 OH2 TIP A 17 22.954 25.044 −0.471 1.0043.52 SS ATOM 2456 OH2 TIP A 18 30.914 51.700 1.620 1.00 43.52 SS ATOM2457 OH2 TIP A 19 18.868 45.761 14.958 1.00 43.52 SS ATOM 2458 OH2 TIP A20 11.192 43.417 6.627 1.00 43.52 SS ATOM 2459 OH2 TIP A 21 21.39533.255 9.085 1.00 43.52 SS ATOM 2460 OH2 TIP A 22 31.566 40.059 −1.4491.00 43.52 SS ATOM 2461 OH2 TIP A 23 26.608 40.843 12.469 1.00 43.52 SSATOM 2462 OH2 TIP A 24 10.882 49.731 3.097 1.00 43.52 SS ATOM 2463 OH2TIP A 25 24.112 6.394 −4.916 1.00 43.52 SS ATOM 2464 OH2 TIP A 26 20.06529.530 9.976 1.00 43.52 SS ATOM 2465 OH2 TIP A 27 26.915 45.385 9.4011.00 43.52 SS ATOM 2466 OH2 TIP A 28 16.232 36.475 12.662 1.00 43.52 SSATOM 2467 OH2 TIP A 29 17.678 25.766 6.317 1.00 43.52 SS ATOM 2468 OH2TIP A 30 20.570 25.957 0.315 1.00 43.52 SS ATOM 2469 OH2 TIP A 31 8.56124.058 9.246 1.00 43.52 SS ATOM 2470 OH2 TIP A 32 20.329 24.123 6.0751.00 43.52 SS ATOM 2471 OH2 TIP A 33 25.927 42.053 15.359 1.00 43.52 SSATOM 2472 OH2 TIP A 34 33.530 38.867 −7.089 1.00 43.52 SS ATOM 2473 OH2TIP A 35 35.591 12.620 10.017 1.00 43.52 SS ATOM 2474 OH2 TIP A 3631.225 41.351 6.010 1.00 43.52 SS ATOM 2475 OH2 TIP A 37 22.123 24.47211.325 1.00 43.52 SS ATOM 2476 OH2 TIP A 38 28.087 29.403 −13.302 1.0043.52 SS ATOM 2477 OH2 TIP A 39 34.560 12.999 5.277 1.00 43.52 SS ATOM2478 OH2 TIP A 40 9.465 48.542 10.702 1.00 43.52 SS ATOM 2479 OH2 TIP A41 12.503 38.149 20.364 1.00 43.52 SS ATOM 2480 OH2 TIP A 42 7.42041.639 10.909 1.00 43.52 SS ATOM 2481 OH2 TIP A 43 35.680 12.866 7.9021.00 43.52 SS ATOM 2482 OH2 TIP A 44 39.502 27.931 18.775 1.00 43.52 SSATOM 2483 OH2 TIP A 45 21.146 25.021 8.714 1.00 43.52 SS ATOM 2484 OH2TIP A 46 30.337 39.275 −10.268 1.00 43.52 SS ATOM 2485 OH2 TIP A 4718.733 12.572 4.205 1.00 43.52 SS ATOM 2486 OH2 TIP A 48 8.456 43.72010.493 1.00 43.52 SS ATOM 2487 OH2 TIP A 49 21.738 21.295 7.389 1.0043.52 SS ATOM 2488 OH2 TIP A 50 20.445 28.671 −16.154 1.00 43.52 SS ATOM2489 OH2 TIP A 51 22.600 12.989 −4.498 1.00 43.52 SS ATOM 2490 OH2 TIP A52 36.376 34.961 13.547 1.00 43.52 SS ATOM 2491 OH2 TIP A 53 42.96428.274 −7.367 1.00 43.52 SS ATOM 2492 OH2 TIP A 54 17.026 16.386 5.4021.00 43.52 SS ATOM 2493 OH2 TIP A 55 20.135 54.487 2.295 1.00 43.52 SSATOM 2494 OH2 TIP A 56 30.869 37.274 6.313 1.00 43.52 SS ATOM 2495 OH2TIP A 57 33.853 40.105 6.671 1.00 43.52 SS ATOM 2496 OH2 TIP A 58 16.34943.334 14.737 1.00 43.52 SS ATOM 2497 OH2 TIP A 59 18.622 23.529 3.1831.00 43.52 SS ATOM 2498 OH2 TIP A 60 26.630 48.674 7.609 1.00 43.52 SSATOM 2499 OH2 TIP A 61 16.555 47.864 −8.705 1.00 43.52 SS ATOM 2500 OH2TIP A 62 23.193 10.211 −8.266 1.00 43.52 SS ATOM 2501 OH2 TIP A 63 6.76133.422 13.017 1.00 43.52 SS ATOM 2502 OH2 TIP A 64 31.235 43.998 6.5741.00 43.52 SS ATOM 2503 OH2 TIP A 65 12.837 27.261 −2.851 1.00 43.52 SSATOM 2504 OH2 TIP A 66 7.112 44.502 1.340 1.00 43.52 SS ATOM 2505 OH2TIP A 67 41.572 21.679 −0.685 1.00 43.52 SS ATOM 2506 OH2 TIP A 6822.680 39.284 19.173 1.00 43.52 SS ATOM 2507 OH2 TIP A 69 39.463 37.601−3.326 1.00 43.52 SS ATOM 2508 OH2 TIP A 70 27.198 44.213 12.911 1.0043.52 SS ATOM 2509 OH2 TIP A 71 20.278 32.081 11.495 1.00 43.52 SS ATOM2510 OH2 TIP A 72 18.798 23.783 14.243 1.00 43.52 SS ATOM 2511 OH2 TIP A73 29.802 13.025 4.755 1.00 43.52 SS ATOM 2512 OH2 TIP A 74 10.47743.339 −5.465 1.00 43.52 SS ATOM 2513 OH2 TIP A 75 36.280 13.938 −3.5061.00 43.52 SS ATOM 2514 OH2 TIP A 76 17.052 11.701 −7.565 1.00 43.52 SSATOM 2515 OH2 TIP A 77 18.133 24.995 −0.393 1.00 43.52 SS ATOM 2516 OH2TIP A 78 38.072 39.296 5.240 1.00 43.52 SS ATOM 2517 OH2 TIP A 79 25.94515.534 −14.313 1.00 43.52 SS ATOM 2518 OH2 TIP A 80 15.465 37.437 19.5491.00 43.52 SS ATOM 2519 OH2 TIP A 81 16.620 20.634 1.922 1.00 43.52 SSATOM 2520 OH2 TIP A 82 23.545 39.853 −13.805 1.00 43.52 SS ATOM 2521 OH2TIP A 83 20.253 35.091 −10.327 1.00 43.52 SS ATOM 2522 OH2 TIP A 8436.698 24.059 13.386 1.00 43.52 SS ATOM 2523 OH2 TIP A 85 13.944 26.7651.654 1.00 43.52 SS ATOM 2524 OH2 TIP A 86 3.518 41.699 1.425 1.00 43.52SS ATOM 2525 OH2 TIP A 87 23.552 46.329 −1.900 1.00 43.52 SS ATOM 2526OH2 TIP A 88 21.467 19.279 −8.361 1.00 43.52 SS ATOM 2527 OH2 TIP A 8911.984 27.072 27.543 1.00 43.52 SS ATOM 2528 OH2 TIP A 90 23.521 11.07923.940 1.00 43.52 SS ATOM 2529 OH2 TIP A 91 28.454 48.510 −3.162 1.0043.52 SS ATOM 2530 OH2 TIP A 92 13.904 43.690 16.624 1.00 43.52 SS ATOM2531 OH2 TIP A 93 7.949 31.840 5.015 1.00 43.52 SS ATOM 2532 OH2 TIP A94 32.500 19.725 14.392 1.00 43.52 SS ATOM 2533 OH2 TIP A 95 38.39028.352 −11.079 1.00 43.52 SS ATOM 2534 OH2 TIP A 96 33.535 30.231 18.6801.00 43.52 SS ATOM 2535 OH2 TIP A 97 42.143 19.333 8.275 1.00 43.52 SSATOM 2536 OH2 TIP A 98 14.934 50.010 −7.142 1.00 43.52 SS ATOM 2537 OH2TIP A 99 30.152 44.543 −0.015 1.00 43.52 SS ATOM 2538 OH2 TIP A 1008.844 47.803 −7.596 1.00 43.52 SS ATOM 2539 OH2 TIP A 101 27.103 37.05119.604 1.00 43.52 SS ATOM 2540 OH2 TIP A 102 17.787 15.577 −9.801 1.0043.52 SS ATOM 2541 OH2 TIP A 104 16.591 25.543 2.635 1.00 43.52 SS ATOM2542 OH2 TIP A 105 27.295 22.133 −0.598 1.00 43.52 SS ATOM 2543 OH2 TIPA 106 8.558 49.822 −12.353 1.00 43.52 SS ATOM 2544 OH2 TIP A 107 6.92243.138 −1.316 1.00 43.52 SS ATOM 2545 OH2 TIP A 108 42.011 13.562 11.6951.00 43.52 SS ATOM 2546 OH2 TIP A 109 20.749 53.219 5.773 1.00 43.52 SSATOM 2547 OH2 TIP A 110 32.142 24.552 22.465 1.00 43.52 SS ATOM 2548 OH2TIP A 111 13.126 28.687 14.559 1.00 43.52 SS ATOM 2549 OH2 TIP A 11236.222 10.792 2.017 1.00 43.52 SS ATOM 2550 OH2 TIP A 113 20.573 36.8561.414 1.00 43.52 SS ATOM 2551 OH2 TIP A 115 27.421 10.974 25.524 1.0043.52 SS ATOM 2552 OH2 TIP A 116 42.751 19.151 10.460 1.00 43.52 SS ATOM2553 OH2 TIP A 117 35.543 36.745 5.730 1.00 43.52 SS ATOM 2554 OH2 TIP A118 11.440 28.831 21.618 1.00 43.52 SS ATOM 2555 OH2 TIP A 119 7.31934.101 9.399 1.00 43.52 SS ATOM 2556 OH2 TIP A 121 36.857 26.359 21.6261.00 43.52 SS ATOM 2557 OH2 TIP A 123 36.523 30.443 7.429 1.00 43.52 SSATOM 2558 OH2 TIP A 124 26.132 50.469 5.525 1.00 43.52 SS ATOM 2559 OH2TIP A 126 34.329 44.402 −4.548 1.00 43.52 SS ATOM 2560 OH2 TIP A 12726.999 33.484 −9.280 1.00 43.52 SS ATOM 2561 OH2 TIP A 128 18.443 27.12421.387 1.00 43.52 SS ATOM 2562 OH2 TIP A 129 17.596 28.119 10.054 1.0043.52 SS ATOM 2563 OH2 TIP A 131 14.334 35.901 18.626 1.00 43.52 SS ATOM2564 OH2 TIP A 132 31.030 32.454 7.688 1.00 43.52 SS ATOM 2565 OH2 TIP A133 2.973 33.533 13.781 1.00 43.52 SS ATOM 2566 OH2 TIP A 137 25.3079.302 −4.123 1.00 43.52 SS ATOM 2567 OH2 TIP A 138 22.308 8.017 22.5391.00 43.52 SS ATOM 2568 OH2 TIP A 140 21.574 38.666 3.205 1.00 43.52 SSATOM 2569 OH2 TIP A 146 31.586 31.424 10.186 1.00 43.52 SS ATOM 2570 OH2TIP A 154 6.490 36.861 3.474 1.00 43.52 SS ATOM 2571 OH2 TIP A 15612.899 47.499 −10.970 1.00 43.52 SS ATOM 2572 OH2 TIP A 160 38.13332.578 21.040 1.00 43.52 SS ATOM 2573 OH2 TIP A 185 16.312 28.123 22.4751.00 43.52 SS ATOM 2574 OH2 TIP A 191 34.865 30.182 −7.396 1.00 43.52 SSATOM 2575 OH2 TIP A 198 38.682 29.942 21.401 1.00 43.52 SS ATOM 2576 OH2TIP A 221 20.664 28.062 −4.027 1.00 43.52 SS ATOM 2577 OH2 TIP A 22738.426 34.659 9.615 1.00 43.52 SS TER END

1. A method of screening potential binding agent(s) to determine those which bind aromatic prenyltransferase(s) wherein said aromatic prenyltransferase(s) have a beta/alpha barrel structure and at least 95% sequence identity with the amino acid sequence set forth in SEQ ID NO:2, said method comprising: contacting said potential binding agent(s) with said aromatic prenyltransferase(s) in the presence of an aromatic prenyltransferase substrate; and determining the ability of said potential binding agent to compete with said aromatic prenyltransferase substrate binding to said aromatic prenyltransferase(s), wherein the potential binding agent has been modeled to fit spatially into an aromatic prenyltransferase interaction site defined by a plurality of atomic coordinates.
 2. A method of identifying a potential substrate of an aromatic prenyltransferase having a beta/alpha barrel structure and at least 95% sequence identity with the amino acid sequence set forth in SEQ ID NO:2, said method comprising: defining an active site of said aromatic prenyltransferase based on a plurality of atomic coordinates of said aromatic prenyltransferase; identifying a potential substrate that fits said active site; and contacting the aromatic prenyltransferase with the potential substrate and determining its activity thereon.
 3. A method of identifying a potential activator of an aromatic prenyltransferase having a beta/alpha barrel structure and at least 95% sequence identity with the amino acid sequence set forth in SEQ ID NO:2, , said method comprising: contacting said aromatic prenyltransferase with a potential activator that fits an active site based on a plurality of atomic coordinates of said aromatic prenyltransferase; and determining the ability of said potential activator to activate said aromatic prenyltransferase.
 4. A method according to claim 1, wherein said aromatic prenyltransferase has the amino acid sequence set forth in SEQ ID NO:2, or conservative variations thereof.
 5. A method according to claim 1, wherein said aromatic prenyltransferase has the amino acid sequence set forth in SEQ ID NO:2.
 6. A method of identifying a potential binding agent for an aromatic prenyltransferase having a beta/alpha barrel structure and at least 95% sequence identity with the amino acid sequence set forth in SEQ ID NO:2, said method comprising: defining an active site of said aromatic prenyltransferase based on a plurality of atomic coordinates of said aromatic prenyltransferase; identifying a potential binding agent that fits said active site; and contacting the aromatic prenyltransferase with the potential binding agent and determining its ability to bind thereto.
 7. A method of screening for compounds that bind to aromatic prenyltransferase(s) having a bet/alpha barrel structure and at least 95% sequence identity with the amino acid sequence set forth in SEQ ID NO:2, said method comprising: testing a compound for the ability to bind to said aromatic prenyltransferase, wherein said compound has been selected as having points of interaction with said aromatic prenyltransferase, and wherein similar points of interaction have been determined between said aromatic prenyltransferase and a substrate or substrate analogue. 